FI68054C - FOERFARANDE FOER FRAMSTAELLNING AV NYA GUANINDERIVAT - Google Patents

Abstract

Novel derivatives of guanine, viz. antiviral compounds of the formula <CHEM> wherein each of R1 and R2, which are the same or different is hydrogen, hydroxy or fluoro; provided that R1 or R2 is hydrogen when R1 and R2 are different, and provided that R1 and R2 are hydroxy or fluoro when R1 and R2 are the same; or a physiologically acceptable salt of an optical isomer thereof, methods for their preparation, pharmaceutical preparations containing the compounds, and methods for the treatment of virus infections and other diseases causes by viruses.

Description

68054

Method for the preparation of new guanine derivatives

The invention relates to a process for the preparation of novel guanine derivatives which can be used for the selective control of viruses, such as herpesviruses, etc., which can cause various diseases in animals, including man. Such diseases include both common infections and tumor diseases, i. cancer.

The effects of viruses on bodily functions are the result of changes that occur at the cellular level and below the cells. Pathogenic changes at the cellular level are different for different combinations of viruses and host cells. Although certain viruses cause certain general cell killing (Killing), others can turn cells into cancer.

Important common viral infections include skin rash (including laryngeal rash), corneal rash, genital rash, shingles, cerebral palsy, infectious glandular fever, and cytomegalovirus infections, all of which are caused by viruses belonging to the herpes virus group. Other important viral diseases are influenza A and B, which are caused by the influenza A and B viruses. Another important common viral disease is viral rupture and especially hepatitis B virus infections are widespread. Effective and selective antiviral agents are needed to treat these diseases as well as other viral diseases.

Numerous different viruses of both DNA and RNA types have been shown to cause cancer in animals. The effect of carcinogenic chemicals in animals can lead to the activation of latent cancer viruses. It is possible that cancer viruses are associated with human cancer. The most plausible 2,68054 cases known to humans today are blood cancers, sarcomas, breast cancers, Burkitt's lymphoid tumors, nasopharyngeal cancers, and cervical cancers with RNA cancers and herpesviruses.

This makes the search for selective inhibitors of cancer-causing viruses and their functions an important task when attempting to treat cancer.

The compound 9- (4-hydroxybutyl) guanine is described in Chem. Pharm. Bull. 17_ (1969) 1268-1270 and Agr. Biol. Chem., 21_ (1973) 2037-2043. However, no antiviral or other pharmacological activity has been demonstrated for said compound.

U.S. Patent No. 4,199,574 discloses a broad group of substituted purines of the formula R1 fVk

In. J-N - CH-X-CH - CH-R

R 2 is N 6 U h R R 6 R 3 R4 wherein X is oxygen or sulfur; R 3 is hydrogen, halogen, hydroxy, alkoxy, azide, thio, alkylthio, amino, alkyl-2-amino or dialkylamine; R is hydrogen, halogen, alkylthio, acylamino, amino or azide; R is hydrogen, straight or branched or cyclic alkyl, hydroxyalkyl, benzyloxyalkyl or phenyl; R is hydrogen, hydroxy or alkyl; R is hydrogen, hydroxy, amino, alkyl, hydroxyalkyl, benzyloxy, benzoyloxy, benzoyloxymethyl, sulfamoyloxy, phosphate carboxypropionyloxy, straight-chain or cyclic acyloxy having 1 to 8 carbon atoms, for example acetoxy or a substituted carbamoyl group of the formula NHCO wherein Z is alkyl, aryl or aralkyl optionally substituted with one or more sulfonyl, aramino, carbamoyl or

It 3 68054 with halogen; is hydrogen or alkyl, provided that when X is oxygen and R 2, R 2, R 1 and R 2 are hydrogen, 1 ex R is amino or methylamino when R is hydrogen or hydroxy. These compounds have been found to have antiviral activity against various classes of DNA and RNA viruses. Examples of particularly active compounds are 9- (2-hydroxyethoxymethyl) guanine and 2-amino-9- (2-hydroxyethoxymethyl) adenine.

The present invention is directed to a compound of formula 0

sK'S

> I>

JJ-N - CH, -CH-CH-CHoOH I

h2n ^^ n ^ l | R 1 R 2 wherein both R 2 and R 2 are the same or different represent hydrogen, hydroxy or fluorine, provided that R 1 or R 2 is hydrogen when R 1 and R 2 are different, and provided that R 1 and R 2 are hydroxy or fluorine when R 1 and R 2 are similar, and to its physiologically acceptable salts or optical isomers.

It has been found that such a compound has antiviral activity and that it inhibits certain viral functions, including tumorigenesis and viral replication.

The invention thus provides a compound and its physiologically acceptable salts, which compounds are useful in the therapeutic and / or prophylactic treatment of viral diseases and which may be useful in the therapeutic and / or prophylactic treatment of viral cancer.

An effective selective antiviral agent, 4 6 8 0 5 4 with acceptable side effects, should have a selective preventive effect against the specific viral function of the virus being fought.

Of particular importance is the use of the compounds of the invention in the treatment of herpes virus infections. Of the herpes viruses, mention may be made of Herpex simplex types 1 and 2, chickenpox virus (shingles), infectious glandular fever virus (i.e. Epstein-Barr virus) and cytomegalovirus. Important diseases caused by herpes viruses include skin rash (including lip rash), genital rash, corneal membrane rash, cerebral palsy, and shingles.

Another potential use for the compounds of the invention is in the treatment of cancer and tumors, especially those caused by viruses. This effect can be achieved in different ways, i. preventing the transformation of virus-infected cells into tumors and preventing the spread of viruses from the altered cells to other normal cells and stopping the growth of cells that have been altered by the viruses.

11 5 68054

Another use for the compounds of the present invention is the inhibition of altered cells due to specific herpesvirus enzymes present in these cells, such as thymidine kinase.

Potential uses of the compounds of the present invention in cancer chemotherapy include the treatment of leukemia, lymphoid tumors, Burkitt's lymphoma and Hodgkins' disease, sarcomas, breast cancer, nasopharyngeal cancers and cervical cancers in which viruses are indicated. Other potential uses of the compounds of the present invention in cancer chemotherapy include multiple bone marrow tumor and lung (as well as bronchial) cancer, stomach, liver, colon, bladder, lips, bones, kidneys, ovary, prostate, pancreas, skin (melanoma), sternum treatment of cancer of the mouth, esophagus, testicles, brain (and cranial spinal cord), thyroid, gallbladder (and ducts), nose, larynx, connective tissue, penis, uterus, vagina, uterine trunk and tongue.

The compounds of formula I may be used to treat diseases caused by A. viruses in animals, including humans, by administering to such an infected animal a therapeutically effective amount of a compound of formula I or a physiologically acceptable salt thereof, B. to prevent multiplication of viruses, in particular herpesviruses, in animals, including humans. in need of such treatment, a compound of formula I or a physiologically acceptable salt thereof in an amount sufficient to prevent said multiplication, C. for the treatment of tumor diseases caused by viruses in animals, including humans, by inhibiting the growth of cells showing viral functions, 6 68054 by therapeutically administering to an infected animal a compound of formula I or a physiologically acceptable salt thereof, D. for inhibiting the growth of virus-modified cells in animals, including humans, by administering to an animal in need of such treatment a compound of formula I or a physiologically acceptable salt thereof in an amount sufficient to prevent said growth, E. to treat tumor diseases caused by viruses in animals, including humans, by inhibiting tumor virus replication, administering to an animal in need of such treatment a compound of formula I or a physiologically acceptable salt thereof. , which is sufficient to inhibit such multiplication, and F. for the treatment of tumor diseases in an animal, including a human, by administering to the animal a therapeutically effective amount of a compound of formula I or a physiologically acceptable salt thereof.

As mentioned above, the compound of the present invention has the formula 0

II

η il "Ί

U '1-n - CH 2 -CH-CH-CH 0 OH I

VR, R2 wherein both and R2, which are the same or different, 78080 denote hydrogen, hydroxy or fluorine, provided that R1 or R2 is hydrogen when R1 and R2 are different, and provided that R1 and R2 denote hydroxy or fluorine , when R 1 and R 2 mean the same, including physiologically acceptable salts and optical isomers.

A preferred subgroup of compounds of the invention is obtained when at least one of R 1 and R 2 in formula I is hydroxy. Another preferred subgroup is obtained when at least one of R 1 and R 2 in formula I is fluorine.

The conditions in the definition of R 1 and R 2 above mean that only the following six specific compounds, including their salts and optical isomers, form part of the present invention: 9- (3,4-dihydroxybutyl) guanine 9- (2,4-dihydroxybutyl) guanine 9- (2,3,4-trihydroxybutyl) guanine (erythro and threo).

9- (3-fluoro-4-hydroxybutyl) guanine 9- (2-fluoro-4-hydroxybutyl) guandine 9- (2,3-difluoro-4-hydroxybutyl) guanine (erythro and threo).

The first compound 9- (3,4-dihydroxybutyl) guanine is a preferred compound of the invention.

The compounds of formula I contain one or two asymmetric centers. Thus, they exist in two or four optical forms, and all such forms are included within the scope of the invention.

methods of Preparation

The compounds of the invention may be prepared by any of the methods A-E below.

β 68054 A. Reduce the compound of formula 0 HN, · Ί,

I '- N - CH - CH (R) CH (R') COOR II

h2n wherein and are as defined above, and wherein R is hydrogen, alkyl of 1 to 8 carbon atoms, substituted or unsubstituted phenyl or benzyl.

The reduction can be carried out with borohydrides, aluminum hydrides, other hydride reducing agents, sodium in ethanol, catalytic hydrogenation or the like by methods known per se for the reduction of esters to alcohols. The reduction is preferably carried out in an organic solvent such as isopropanol with metal borohydrides, dioxane with aluminum hydrides or ethanol in catalytic hydrogenations. The reaction temperature is preferably 0 ° to 50 ° C for 1 hour to 3 days.

B1. The compound of formula Y is hydrolyzed

. J - .Nx n | Ί

i 1 _________N - CH0CH - CH - CH „OR5 IV

HN '"-N" · _ I 1 1 ^ 6 H OR3 4

OR H

4 3

OR OR

H F

F H

F F

wherein Y is OH or a group which can be converted by hydrolysis to a hydroxy group such as chlorine, bromine, iodine or a group -SH, 2 2 2 2 -OR, -SR or -SO 2 R, wherein R is alkyl of 1 to 8 carbon atoms, 1- Fluorinated alkyl or alkylaryl having 8 carbon atoms, such as benzyl or aryl, and R 1, R 2, R 1 * and R 2 2 are the same or different and represent hydrogen, -COR or -SO 2 R, where R 1 is as defined above.

The reaction is preferably carried out in water with an acid or base such as hydrogen chloride or sodium hydroxide, at a temperature of 20 ° and 100 ° C for 1-24 h.

Il 9 68054

B2. Hydrolyse of formula IV as determined in Method B1

wherein R 3 and R 4 together or R 3 and R 5 4.5 together or R 3a R together form a cyclic ketal as defined in Method E or a cyclic carbonate ester.

The reaction conditions for hydrolysis are described in Method B1.

C. Condensing a compound of formula R7 N t

I 'I

with a compound of formula X2 - CH ~ - CH - CH - CH "R1 °" h r8-

R9 H

R9 R8

H F

F H

F F- 7 wherein R is a silyloxy group such as trimethylsilyloxy or tert. - butyldiphenylsilyloxy or a group Y as defined in method B1, R is a silyl group such as trimethylsilyl or tert-butyldiphenylsilyl or a group R6 as defined in method B1, and R, R and R are the same or different and denote silyl groups such as trimethylsilyloxy or tert-butyldiphenylsilyloxy 1 2 3 or groups Z, Z 7a Z which are identical or different and denote halogen such as chlorine, bromine or 3 4 5 iodine, or groups OR, OR or OR as defined in Method 2 or Method B2, and X must remove a group 2 such as chlorine, bromine, iodine or a group SO 2 R, where R is as defined in Method UI, followed by hydrolysis of the groups R 1 -R 2. .

68054 10

The condensation is preferably carried out in an organic solvent such as dimethylformamide or ethanol and at a temperature between 0 ° and 100 ° C for 1 hour and 3 days in the presence of a base such as potassium carbonate.

In addition to using the reaction conditions for hydrolysis described in Method B1, some silyl groups, such as trimethylsilyloxy, can be further hydrolyzed in a solvent such as water or methanol, and other silyl groups such as tert-butyldiphenylsilyloxy can also be hydrolyzed by treatment with fluoride ion such as ° and 50 ° C for 1-24 h.

D. Ring a compound of formula R7 '-r nh2, n

IIXT S — NH - CH "- CH - CH - CH" R1 ° VII

HN 2 i i 2 i "'r8>

H H

R8 R8 F F H H _F f J

7 8 9 10 11 wherein R, R, R, R and R have the same meaning as in process C, by reacting it with formic acid or a derivative thereof such as orthoformate ester and then hydrolyzing the groups R -R thereafter.

The ring closure is preferably performed in an organic solvent such as formamide or ethanol and at a temperature between 20 ° and 210 ° C for 1-10 h.

n 68054 1 1 E. Condensing a compound of formula R7 N · ^ | i.! I xi1 NH with a compound of formula k "X2-CHO-CH-CH-CH_ or X2-CH -CH-CH-CH-or X2-CH-CH-CH-ai OR5 2 ^ | I 2 2 il \ λ 2 t | 2

0 0 O O O O

V V XI11 R1 'V2 R1' XR12 »'' V2 1 12 wherein R is as defined in Method A, R is alkoxy of 1 to 8 17 11 2 carbon atoms or R; R, R and X are as defined in Method C and R 1 is as defined in Method B1, after which the protecting groups are hydrolysed to form a compound of formula I wherein R 1 or R 2 is hydroxy.

The condensation reaction conditions are described in Method C.

The methods A-E described can be used to provide mixtures of enantiomers or, where appropriate, to provide a single enantiomeric isomer. In addition, a single enantiomeric isomer can be obtained from enantiomeric mixtures by methods known per se.

The starting materials in the above methods A-E are either known compounds or can be prepared by known methods.

salts

Physiologically acceptable salts of the compounds of the invention are prepared by methods known per se below. Salts are new compounds. Metal salts can be prepared by reacting a metal hydroxide with a compound of the invention. Examples of metal salts that can be prepared in this way include salts containing lithium, sodium and potassium. The less soluble metal salt can be precipitated from the more soluble salt solution by adding a suitable metal compound. Acid salts can be prepared by reacting a compound of the invention with an acid such as HCl, HBr, SO 2 or organic sulfonic acid.

Pharmaceutical preparations

In clinical use, the compounds of the invention are usually administered topically, orally, nasally or by injection or inhalation in the form of a pharmaceutical preparation containing the active ingredient in the form of the parent compound or optionally a pharmaceutically acceptable salt thereof together with a pharmaceutically acceptable carrier which may be a solid, semi-solid or liquid diluent. or a digestible capsule, and such preparations form a further feature of the invention. The compound can also be used without a carrier. Examples of pharmaceutical preparations include tablets, drops such as nasal drops, eye drops, topical preparations such as creams, jellies, ointments and suspensions, inhalable sprays, nasal sprays, liposomes, etc. The active ingredient usually contains 0.01 and 99 or 0.1 and 99 parts by weight. -% of the preparation, for example 0.5 and 20% in preparations intended for injection and 0.1 and 50% in preparations intended for oral use.

The preparations are preferably in unit dosage form. They are furthermore preferably available in sterile form.

To prepare pharmaceutical preparations in the form of oral dosage units containing a compound of the invention, the active ingredient may be mixed with a solid, powdered carrier such as lactose, sucrose, sorbitol, mannitol, starch such as potato starch, corn starch or corn starch, amylopectin, laminate may also contain lubricants such as magnesium or calcium stearate 13 68054 or carbowax or other polyethylene glycol waxes and may be compressed to form tablets or granules. If granules are desired, the cores can be coated, for example, with concentrated sugar solutions which may contain gum arabic, talc and / or titanium dioxide or, alternatively, with a film-forming agent dissolved in volatile organic solvents or mixtures of organic solvents. Dyes can be added to these coatings, for example to separate the different contents of the active substance. To prepare soft gelatin capsules consisting of gelatin and, for example, glycerol and a plasticizer, or to prepare such sealed capsules, the active ingredient may be mixed with a carboxylic wax or a suitable oil such as sesame oil, olive oil or arachis oil.

Hard gelatin capsules may contain granules of the active ingredient with solid, powdered carriers such as lactose, sucrose, sorbitol, mannitol, starches (for example potato starch, corn starch or amylopectin), ointment as cellulose derivatives or gelatin, and may also contain magnesium stearate, and may also contain magnesium

By using the active drug in several layers separated by slowly dissolving coatings, sustained release tablets are obtained. Another way to prepare slowly dissolving tablets is to divide the dose of active drug into several granules coated with coatings of different thicknesses and compress the granules into tablets together with a carrier. The active substance can also be added to slowly dissolving tablets prepared, for example, from fatty and waxy substances or evenly distributed in a tablet of an insoluble substance, such as a tablet of a physiologically inert plastic substance.

Oral liquid preparations may be in the form of elixirs, syrups or suspensions, for example solutions containing from about 0.1% to about 20% by weight of active ingredient, sugar and ethanol, water, glycerin, propylene glycol and optionally flavorings, saccharin. and / or solutions of carboxymethylcellulose as dispersants.

Formulations for parenteral injection may contain an aqueous solution of the active ingredient or a physiologically acceptable salt thereof, preferably in a concentration of 0.05-10%, and optionally also a stabilizer and / or buffers in aqueous solution. The dosage units of the solution can advantageously be enclosed in ampoules.

For external administration, in particular for the treatment of herpesvirus infections of the skin, genitals and mouth and eyes, the preparations are preferably in the form of a solution, ointment, gel, suspension, ointment or the like. The amount of active substance may vary, for example between 0.05 and 20% by weight of active substance. Such topical preparations can be prepared in a known manner by mixing the active ingredient with known carriers such as isopropanol, glycerin, paraffin, stearyl alcohol, polyethylene glycol, and the like. The pharmaceutically acceptable carrier may also contain a known chemical absorption enhancer. Examples of absorption enhancers are, for example, dimethylacetamide (U.S. Pat. No. 3,472,931), trichloroethanol or trifluoroethanol (U.S. Pat. No. 3,391,757), certain alcohols and mixtures thereof (British Patent 1,001,949).

The dose at which the active ingredients are administered may vary over a wide range and depends on various factors such as the severity of the infection, the age of the patient, etc., and may need to be adjusted individually. As a possible range for the amount of the compounds of the invention that can be administered daily, mention may be made of about 0.1 mg to 2000 mg or about 1 to 2000 mg or preferably 1 to 2000 mg for topical administration, 50 to 2000 mg or 100 to 1000 mg for oral administration and 10 to 2000 mg or 50 to 500 mg when injected.

In severe cases, it may be necessary to increase these doses fifty-fold. In less severe cases, the use of 500 to 1000 mg may be sufficient.

Pharmaceutical compositions containing the active ingredients are preferably formulated so as to provide dosages in these ranges, either as single dosage units or as multiple dosage units.

According to the invention, it has thus been found that the compounds of the formula I and their physiologically tolerable salts can be used for the prevention of the growth of the herpes virus. The compounds of the formula I and their physiologically tolerable salts are useful in the therapeutic and / or prophylactic treatment of viral infections.

The use of the compounds of the formula I or their physiologically tolerable salts in the treatment of herpesvirus infections is a preferred embodiment of the invention.

EXAMPLES

The preparation of the compounds of the invention is illustrated by the following examples.

I. Manufacture of starting materials

Example 1. Preparation of 4-bromo-2-hydroxybutyl acid ethyl ester

Br-CH ~ CHoCH-C00CHoCH,

OH

1 S · 16 68054 2-hydroxybutyrolactone, British Patent Publication 688,253 / C. A. 48, p.3996 (1954) (5.1 g) was dissolved in 10 ml of ethanol and the solution was saturated with hydrogen bromide at 0. After standing at room temperature for 66 h, the solvent was evaporated under reduced pressure. and the mixture was neutralized with 10% aqueous sodium carbonate solution, the mixture was then extracted several times with diethyl ether, and the combined ether extracts were washed with saturated aqueous sodium sulfate solution and dried over anhydrous sodium sulfate, and the residue was distilled off at 12 mm Hg.

In Example 2 below, a fraction boiling at 109-112 ° C and weighing 3.79 g was used.

Example 2. Preparation of 4- (2-amino-6-chloropurin-9-yl) -2-hydroxy-butyric acid ethyl ester

Cl

-OF

l! l JJ

Η ,, Ν · ^ N ^ V N OH

2W I | CH 2 CH 2 CH-COOCH 2 CH 3 2-amino-6-chloropurine (0.509 g, 3.00 mmol), 4-bromo-2-hydroxybutyric acid ethyl ester (0.633 g, 3.00 mmol; prepared according to Example 1) and anhydrous potassium carbonate (0.415 g, 3.00 mmol) was mixed with 10 ml of dimethylformamide and the mixture was stirred at room temperature for 65 h. The mixture was then filtered and the filtrate was evaporated at 0.1 mm Hg. The crystalline residue was triturated with 8 ml of chloroform, and the insoluble matter was removed by filtration and washed with 2 ml of chloroform. The resulting material was then triturated with 5 ml of water, and the insoluble matter was removed by filtration and washed with 2 ml of water. Refilling with 11 ml of ethanol gave 0.360 g of product. Sp. 163-164 ° C (uncorrected) 17 6 8 0 5 4 UV spectrum (ethanol): λ (nm) 311, 248. Analysis - found: C 43.90; H 4.78; Cl 11.72; N 23.52; O 15.90%. Calculated for C 11 H 14 ClN 5 O 3: C, 44.08; H 4.71; Cl 11.83; N 23.37; O 16.01%.

Example 3. Preparation of 4- (2-amino-1,6-dihydro-6-oxopurin-9-yl) -2-hydroxybutyric acid__

O

A

HN ^ A-N

I 1 I)

H ν'Ά'Ά ^ OH 2 I I

CH2CH2CH-COOH

4- (2-Amino-6-chloro-purin-9-yl) -2-hydroxybutyric acid ethyl ester (1.40 g, 4.67 mmol; prepared according to Example 2) in 100 mL of 1M aqueous hydrochloric acid was refluxed at 2.5 h. The solvent was then evaporated at a pressure of about 10 mm Hg. Water (25 ml) was added to the residue and the solvent was evaporated again. This delivery was delivered 4 times. The residue was triturated with 150 ml of acetone and the semi-solid was removed by filtration to give 1.38 g. This crude product (1.27 g) was partially dissolved in 5 ml of water, the solution was filtered and the insoluble matter was washed with 2.5 ml of water. The pH of the filtrate was then adjusted to 6-7 with solid sodium bicarbonate. The resulting aqueous solution was filtered, and the insoluble matter was washed with 7.5 ml of water. Acetic acid was then added to 0.4 ml of the filtrate. After cooling to 0 ° C, the precipitate was removed by filtration, washed with 3 ml of water. Recrystallization from 75 ml of water gave 0.68 g of product. Sp. > 200 ° C (decomposes). UV spectrum (0.1M hydrochloric acid): λmax (nm) 279, 254; UV spectrum (0.1M sodium hydroxide): λmax.s (nm) 269, 256 (inf.).

Analysis found: C 42.63; H 4.41; N 27.74; O 25.30%. Calculated for C 9 H 11 N 5 O 4: C 42.69; H 4.38; N 27.66; 0 25.27.

ib 6 8 0 5 4

Example 4. Preparation of 4- (2-amino-1,6-dihydro-6-oxopurin-9-yl) -2-hydroxybutyric acid ethyl ester

O

i i] 1

^ ^ N

CH2CH2JHCOOC2H5 4- (2-amino-1,6-dihydro-6-oxopurin-9-yl) -2-hydroxybutyric acid (2.00 g, 7.9 mmol; prepared according to Example 3) was stirred with 500 mL ethanol. The mixture was saturated with hydrogen chloride gas, first without cooling and then under ice-water cooling. The total addition time was about 15 min. The mixture was then slowly warmed to room temperature and allowed to stand overnight. After evaporation of the solvent, the residue was treated three times with 25 ml of ethanol each time, the solvent being evaporated again after each treatment. The residue was then dissolved in 12 ml of water and the pH was adjusted to 6-7 with saturated aqueous sodium bicarbonate solution. The precipitate was removed by filtration, washed with 2 ml of water and dried in vacuo to give 1.60 g. Recrystallization from ethanol gave the pure product, m.p. 161-163 ° C (the analytical sample had a melting point of 162-163 ° C). Analysis found: C 46.96; H 5.35; N 24.77; O 22.60%. Calculated for C 11 H 15 [5N 5 O 4: c 46.97; H 5.38; N 24.90; O 22.74%.

Example 5. Preparation of 4- (2-amino-6-chloropurin-9-yl) methyl-1,3- __-dioxane

C

L .N

ΓΥ1 H-N /

// CH CU

ch9-chC "O

^ ^ O CH2X

19 680 5 4

Equimolar amounts of 2-amino-6-chloropurine, 4-bromomethyl-1,3-dioxane (prepared according to Price C.C. J.Amer.

Chem. Soc. 1950, 72, 5335-6) and anhydrous potassium carbonate was mixed with dry dimethylformamide. After stirring for 7 days at room temperature, the mixture was filtered and the filtrate was evaporated under reduced pressure. The residue was triturated with hot ethanol and the insoluble matter was removed by filtration. The filtrate was evaporated to dryness to give a yellow crystalline solid which was recrystallized from chloroform to give colorless rod-shaped crystals. Sp. 169-70 ° C (uncorrected) UV spectrum (hydrochloric acid 0.01 mol / l); 308, 245. UV spectrum (ethanol): (nm) 310, 247. MS: 11.2 a J., m / e (int.): 271/269 (0.19 / 0.55), 226 / 224 (0.11 / 0.28) 212/210 (0.33 / 0.82) 171/169 (0.35 / 1.0).

Example 6. Preparation of 2-fluorobutyrolactone 2-Bromobutyrolactone (15 g) and silver fluoride (23 g) were stirred for 24 hours in refluxing dry acetonitrile (150 ml). After freezing to room temperature, the mixture was filtered and the solvent was evaporated in vacuo. The dark residue was dissolved in ethyl acetate (200 mL) and washed with water (6 x 50 mL). The aqueous phase was backwashed with ethyl acetate (80 ml) and the combined ethyl acetate solution was dried over magnesium sulphate and evaporated to give 2-fluorobutyrolactone (21%). 1 H-NMR (acetone-d 6, Me 2 Si): δ 5.02 and 5.95 with CHFr (two triplets, J = 8 Hz. J 1 = 51 Hz).

Example 7. Preparation of 4-bromo-2-fluorobutyric acid ethyl ester Hydrogen bromide gas was added over about 30 minutes to an ethanolic solution of 2-fluorobutyrolactone (1.8 g) (15 ml) at 0 ° C. The reaction mixture was left at room temperature for 3 days, after which it was evaporated in vacuo. The residue was dissolved in ice-water, the solution was neutralized with sodium carbonate and extracted with ether. The ether solution was washed with saturated aqueous sodium sulfate, dried over sodium sulfate and evaporated to give 4-bromo-2-fluorobutyric acid ethyl ester (1.26 g, 34%). nj = 1.4698.

68054 1 K-NMR (COCl 3, MeSi) δ: 1.3 (t, J = 7Hz, CH 3); 2.1-2.8 (m, CH 2); 3.55 (t, J = 7Hz, CH 2 Br); 4.3 (q, J = 7 Hz, CH 2); 4.75 and 5.55 (two triplets, J = 6Hz, CHF, J = 50 Hz).

LGB

Example 8. Preparation of 4- (2-amino-6-chloropurin-9-yl) -2-fluoro-butyric acid ethyl ester

Cl xx> H N N 2 · CH2CH2CH-CO2CH2CH3

F

4-Bromo-2-fluorobutyric acid ethyl ester (0.23 g), 2-amino-6-chloropurine (0.19 g) and anhydrous potassium carbonate (0.15 g) were stirred in dimethylformamide (3.7 ml) at room temperature for 2 days. The solution was filtered and evaporated in vacuo. The residue was triturated with chloroform (4 + 2 ml) and the clear chloroform solution was evaporated to give the desired product (0.24 g, 72%). TLC (silica gel, i-propanol-water-concentrated ammonia 8-1-1): Rf = 0.76. NMR (CDCl 3, Me 2 Si) δ: 1.26 (t, CH 3); 2.4-2.5 (m, CK2); 4.2-4.3 (t and q, NCH 2 and CO 2 CH 2); 4.77 and 5.03 (CHF, J = 5.1 Hz); 5.2 (NH2); 7.77 (8-CH).

Example 9. Preparation of 4- (2-amino-6-chloropurin-9-yl) -2-fluorobutan-___nol __________________________

u M N N

H2N I

CH2CH2CHCH20H

F

Sodium borohydride (15 mg) was dissolved in dry diethylene glycol diethyl ether (0.9 ml) and stirred. Finely ground and dry lithium bromide (34 mg) was added, followed after 1/2 hour by the addition of 4- (2-amino-6-chloro-purin-9-yl) -2-fluorobutyric acid ethyl ester (100 mg). mg). The Rcak thio mixture was heated at 100 ° C for 3 hours, then poured onto crushed ice (10 g) supplemented with concentrated hydrochloric acid (0.5 ml) and stirred. The pH was adjusted to 6.5 with sodium hydrogen carbonate, the solution was evaporated in vacuo and the residue was redissolved in methanol-chloroform (40-60, 10 ml) and filtered. The solution was purified by chromatography on a silica gel column eluting with a methanol-chloroform gradient to give the desired product (52 mg, 60%) TLC (silica gel, methanol-chloroform 40-60): Rf -0.66-UV (ethanol) Amakg: 310 and 247 nm.

Example 10. (E) -4- (2-Amino-6-benzyloxypurin-9-yl) - ____ 1,2-O-isopropylidene-1,2-dihydroxy CH 2 Cl 2 H 5

Xx> h2n n j CH »-CH -— CH -CH» (S) 2 2 i | 2

O O

X

CH3 ch3 (S) -1,2-O-isopropylidenebutane-1,2,4-triol (Preparation of (S) - (-) - maleic acid according to Hayashi H. et al., J. Amer.

Chem. Soc. 1973, 92, 8749-8757) were mesylated (according to the method of Crossland R. <. And Servis KL, Org.Chem. 1970, 35, 3195) and (S) -4-O-methanesulfonyl-1,2-O-isopropylidenebutane 1,2,4-Triol was used without further purification.

2-Amino-6-benzyloxypurine (Bowles WA et al. Med. Chem. 1963, 6, 471-480) and tetrabutylammonium hydroxide in water were extracted with chloroform and the extract was dried (MgSO 4) and evaporated to give 2-amino-6-benzyloxypurine. benzyloxy-puriinitetrabutyyliammoniumsuolaa.

22 68054 2-Amino-6-benzyloxypurine tetrabutylammonium salt (300 mg, 0.62 mmol), (S) -4-O-methanesulfonyl-1,2-O-isopropylidenebutane-1,2,4-triol (140 mg, 0, 62 mmol), and a chloroform solution of tetrabutylammonium iodide (22 mg, 0.06 mmol) (10 mL) was refluxed for 24 h and then evaporated in vacuo. Chromatography (silica gel, methanol-chloroform 5-95 eluent) gave 72 mg (31% yield) of the title compound.

After treatment with water, vacuum drying and treatment with small portions of ether, an off-white solid appeared as a single spot on TLC (silica gel, CHCl 3 -MeOH 20-1, Rf 0.33). 1 H N MR (CDCl 3): 1.36 and 1.44 (2s, 2x3 H) isopropylidene group; 2.1 (m, 2 H) 2 H 2; 3.6-4.4 (m, 5H) 2Ηχ, 1H3, 2H4; 5.15 (broad s, 2H) NH 2; 5.64 (s, 2H) benzylic CH 2; 7.5 (m, 5H) phenyl; 7.73 (s, 1 H) Hg.

II. Preparation of the compounds of the invention

Example 11. Preparation of 9- (3,4-dihydroxybutyl) guanine 1¾ H2N N / / CH2-CH2-CH-Cl2OH (VIS 707)

OH

To a suspension of ethyl 4- (2-amino-1,6-dihydro-6-oxopurin-9-yl) -2-hydroxybutyrate (prepared according to Example 4) in isopropanol was added excess sodium borohydride and the mixture was refluxed overnight (at least 8 hours). . Hydrochloric acid was added until a clear solution (neutral pH) was obtained. After removal of the solvent, the residue was dissolved in as little boiling water as possible and kept at 0 ° C for a couple of hours. The solid was removed by filtration. The filtrate was evaporated under reduced pressure and the residue was dissolved in hydrochloric acid (1 mol / l) and adsorbed onto a cation exchange resin (Dowex 50 W, H + form). The resin was washed with water and then eluted with 5% ammonium hydroxide. The eluent was evaporated to give a crystalline solid which crystallized from water to give colorless needles. Sp. 260-1 ° C (decomposes) (uncorrected) UV spectrum (hydrochloric acid 0.01 mol / l): λ (nm) 277, 253 (c. = 11500) MS: 11.2 a J. (int.): 239 (M +, 0.13), 222 (0.19), 221 (0.11), 152 (0.43), 151 (0.56), 44 (1.0).

Example 12. Preparation of 9- (2,4-dihydroxybutyl) guanine

O

Jk U_-Nl (VIS 715) H2N /

CH 2 = H-CH2CH2OH

OH

4- (2-Amino-6-chloropurin-9-yl) methyl-1,3-dioxane (prepared according to Example 5) was dissolved in hydrochloric acid (1 mol / l) and refluxed for 4 hours. The solution was basified with dilute ammonium hydroxide and evaporated to dryness under reduced pressure. The residue was dissolved in water and purified by preparative HPLC on a reverse phase column (yU Bondapack C ^ g), eluting with a mixture of methanol and water (1: 3). The product obtained was a white crystalline solid. Sp. 226-8 ° C (decomposes) (uncorrected) UV spectrum (hydrochloric acid 0.01 mol / l): Amaks (nm) 277, 254 (c = 107O0).

Example 13. Preparation of 9- (3-fluoro-4-hydroxybutyl) -guanine __ H, N (VIS 912)

H N ™ V F

2 I I

CH CH2CHCH OH

24 68054 4- (2-Amino-6-chloropurin-9-yl) -2-fluorobutanol (52 mg) was heated in refluxing aqueous 1M HCl (4.5 mL) for 50 minutes. The solution was evaporated in vacuo, the residue was dissolved in blood and the solution was neutralized with 1M ammonia and lyophilized. The residue was purified on an anion exchange column (Dowex 1x2, OH). The evaporated solution gave the desired product (37 mg, 77%).

NMR (DMSO-d 6) δ: 1.95-2.15 (m, CH 2); 3.3-3.5 (m, CH 2); b 2 2 4.05 (t, NCH 2); 4.3 and 4.5 (CHF, J = 49 Hz); 6.45 (NH2); 7.7 (8-CH); 10.6 (NH).

Example 14. Preparation of 9- (DL-erythro-2,3,4-trihydroxybutyl) - __ guanine_______________ Λλ HN, 1 L (VIS 925)

y · N-CH_-CH-CH-CHo0H

H, N N 2 I I 2

OH OH

6-Chloroguanine (0.75 g) and sodium hydride (60% in oil, 0.18 g) in dry dimethylformamide were stirred at room temperature for 1 hour. DL-erythro-2,2-dimethyl-4-p-toluenesulfonyloxymethyl-5-hydroxymethyl-1,3-dioxoacids / 0.70 g, prepared according to A. Holy, Coll. Czech.Chem. Commun Vol 44, pages 593-612 (1979) was added and the reaction mixture was heated at 100 ° C for 17 hours, then filtered through a pad of Celite and the solvent evaporated in vacuo. The residue was refluxed with acetic acid (25 ml) for 2 hours. evaporated and the residue co-evaporated with water (3x5 ml) The residue was dissolved in a small volume of 1N hydrochloric acid and applied to a Dowex 50x8 column (ti ', 70 ml) eluted with water (1 liter) followed by aqueous ammonia (diluted 1 Elution was monitored with a UV detector, the appropriate fractions were evaporated in vacuo and the residue co-evaporated with water several times, finally dissolved in boiling water (230 ml) and filtered to give 9- (DL-ery tro-2.3 , 4-trihydro-di-butyl i) guanine (0.20 g) NMR (DMSO-d 6): 7.8 (s, 8C-II), 6.4 (NH 2), 4.22 (N- CH2).

Example 15 15. Preparation of (S) -9- (3,4-dihydroxybutyl) -guanine___ _______________________

O

HtA, .- '' i 11 N7

H2M I

CH „.CHnCH-CH„ 011 (S) Z SL ’Z -

OH

Treatment of (S) -4- (2-amino-6-benzyloxynurin-9-yl) -1,2-O-ipropyl-diene-1,2-hydroxybutane with hydrochloric acid) 1 mol / l) At 100 ° C for 1 hour gave (S) -9- (3,4-ethylhydrobutyl) guanine.

TLC (silica gel, isopropanol-water-concentrated ammonia 7: 2: 1, 0.60), HPLC Bondapak Phenyl / porasil B column, methanol-water 30-70), and UV (hydrochloric acid 0.1 mol / l) were identical with a racemic compound (VIS 707).

Example 16. Preparation of R- (+) - 9- (3,4-dihydroxybutyl) guan _________preparation_____

Step a: Preparation of R - (+) - 1,2,4-butanetriol R- (+) - dimethyl melt (1.62 g, 10 mmol) which was. prepared by Bogerin, D.L. and Panekin, J.S. according to J. Org. Chem. 19b ;, s 6, 1208-10, was dissolved in THF (10 mL) and added dropwise to a preheated suspension of lithium aluminum hydride 5 · (0.63 g, 16.5 mmol) in THF (15 mL). The reaction mixture is stirred; overnight. 55 ° C. After successive addition of ety verp; (0.62 ml), 10% sodium hydroxide dia (1.20 ml) and water Π 9 n; the mixture was filtered and the solid residue was boiled twice; > (2 x 20 mL.) and filtered. The filtrates were combined with evaporation under reduced pressure (13 Pa, 30 ° C) to give ...

26 68054 crude 1,2,4-butanetriol (0.7 g, 6.6 mmol), 66%.

Step b: Preparation of R - (+) - isopropylidene-Butane-1,2,4-triol R - (+) - Butanetriol (0.7 g, 6.6 mmol) prepared as described in step a was stirred for 1 h. For 5 hours in acetone (50 ml) containing 3 drops of concentrated percarbonate, saturated aqueous sodium bicarbonate solution (2 ml) was added and stirring was continued for a further 10 minutes. The precipitate was filtered off and the filtrate was evaporated under reduced pressure (2.7 Kpa, 30 ° C). The residue was dissolved in ethyl acetate, washed with saturated aqueous sodium bicarbonate solution (5 ml) and brine (5 ml), and dried over magnesium sulfate. The solvent was evaporated and the residue was distilled to give the title compound as a colorless oil (0.3 g, 2.05 mmol), mp 104-6 ° C at about 2.7 kPar, n = 1.4390.

Step c: Preparation of R- (+) - 4-bromoisopropylidene.butane-1,2-diol with R - (+) - isopropylidene-butane-1,2,4-triol (0.3 g, 2.05 mmol ) and triphenylphosphine (0.63 g, 2.4 mmol) were dissolved in methylene chloride (5 mL) and frozen at 0 ° C. N-Bromosuccinimide (0.38 g, 2.16 mmol) was added in small portions with stirring at 0 ° C. After stirring for an additional 1 h at 0 ° C, hexane (15 mL) was added and the resulting precipitate was removed by filtration and washed twice with hexane (2 x 5 mL).

The combined hexane solutions were passed through a short silica gel column (5 g). Elution with hexane (15 ml) after evaporation and distillation gave the title compound as a colorless oil (0.2 g, 0.96 mmol, 47?): Mp 20-20 74-6 at 2.7 kPa (20 mm Hg). , n ^ = 1.4630. + - 27.7 (c = 20, CHCl 3).

Step d: Preparation of R - (+) - 4- (2-amino-6-chloropurin-9-yl) -isopropylidene-butane-1,2-diol from 2-amino-6-chloropurine (162 mg, 0, 96 mmol), R - (+) - 4-bromoisopropylidene-butanediol (200 mg, 0.96 mmol) and potassium carbonate (132 mg) were stirred in DMF (10 mL). After stirring for 16 hours at 68054, the reaction mixture was filtered through celite and the solvent was evaporated under reduced pressure (13 Pa (0.1 mm Hg), 50 ° C). The residue was triturated with warm chloroform (5 ml), and the insoluble matter was filtered off.

The solvent was evaporated to give a pale yellow crystalline solid consisting mainly of the 9 and 7 isomers. These were separated by silica gel flash chromatography. Elution with chloroform / methanol (15: 1) gave the title compound in pure form (106 mg, 0.36 mmol, 37%): mp 12-30 ° C, = + 57.5 (c - 6.97, CHCl 3). j).

Step e: Preparation of R - (+) - 9- (3,4-dihydroxybutyl) guanine

The R - (+) - 4- (2-amino-6-chloropurin-9-yl) isopropylidene-butane-1,2-diol (100 mg, 0.33 mmol) prepared according to step d was dissolved in hydrochloric acid (1 mol). / 1) and refluxed for 1 hour. The solution was concentrated in vacuo and the residue was dissolved in water (5 mL) and made alkaline by the addition of aqueous ammonium hydroxide. After evaporation, the solid residue was recrystallized from water to give the title compound as white needles (40 mg, 0.17 mmol, 51%). UV spectrum (hydrochloric acid 0.01 mol / l): (nm) 277, 253. [α] D = + 30.8 (c = 0.25, water). PMR (DoO max * to internal ref. T-BuOH, δ = 1.23 ppm), 1.75-2.15 ppm (m, 2H), 3.40-3.70 ppm (m, 3H), 4.1 ppm (AB, 2H) and 7.77 ppm (s, 1H).

q

Example 17. R- (+) - 9- (3,4-dihydroxybutyl) guane. Preparation of i-hydrochloride (R) -9- (3,4-dihydroxybutyl) guanine (5.02 g, 21.0 mmol) was dissolved in 5 M hydrochloric acid (5.6 mL, 28 mmol) with heating. After crystallization at 0 ° C, the mixture was lyophilized to give the hydrochloride in quantitative yield, melting point about 160 ° C (after vacuum drying at 110 ° C).

1 H NMR (D 2 O): 61.9-2.3 (m, 2H) NCH 2 CH 2; 3.5-3.9 (m, 3H) CHCl 3; 4.39 (t, 2H) NCH; 4.9 (s) HDO; 8.91 (s, 1H9 H0).

/ O

28 68054

Example 18. Preparation of 9- (3,4-dihydroxybutyl) guanine

Step a: Preparation of 4- (2-amino-5-nitro-6-oxy-pyrimidin-4-yl) -1,2-O-isopropylidene-1,2-butanediolamine

O

H x N0?

NN

H2N'X ^ V ^ N ^ XNHCH2ai2ai _CH2

I I

o o

V

H 3 C ch 2-amino-4-chloro-5-nitro-6-pyrimidinol (3.80 g, 20 mmol), potassium bicarbonate (2.00 g, 20 mmol) and 1-amino-3,4-O-isopropylidene- dihydroxybutane (2.92 g, 20 mmol) in 40 mL of dimethylformamide was stirred overnight. The solvent was evaporated and the residue was triturated with 50 ml of ethanol. The white solid was removed by filtration, dried and recrystallized from 1 L of ethanol. Insoluble material 2.29 g was removed by filtration. 1.88 g of white crystals were removed from the ethanol solution by filtration.

Step b: Preparation of 9- (3,4-dihydroxybutyl) guanine 4- (2-amino-5-nitro-6-oxo-pyrimidin-4-yl) -1,2-O-isopropylidene-1,2- butanedioliamine (283 mg, 1 mmol) in 100 mL of ethanol was hydrogenated using 5% palladium on charcoal at 4 atm for 72 hours. The solvent was evaporated under nitrogen and diethoxymethyl acetate (5 mL, 31 mmol) was added.

The mixture was allowed to stand overnight. Excess diethoxymethyl acetate was evaporated and trifluoroacetic acid (5 mL, 65 mmol) was added. The mixture was allowed to stand in the refrigerator for 5 days. The product was analyzed by HPLC using 30? ·. methanol in water on a g-column and the product was identical to the compound prepared according to Method A (Example 11). HPLC shows more than 88% yield.

29 6 8 0 5 4

Example 19. Preparation of (R) -9- (3,4-dihydroxybutyl) guanine d-camphorsulfonate _________

A mixture of (R) -9- (3,4-dihydroxybutyl) guanine (0.50 g, 2.09 mmol) and d-camphorsulfonic acid (0.50 g, 2.15 mmol) was dissolved with heating at 1.5 ml of 2-propanol, cooled and stirred to accelerate crystallization and filtered to give 0.896 g (80%) of the title compound. Recrystallization from 95% aqueous 1-propanol and ethyl acetate gave a melting point of 190-2 ° C.

1 H NMR (D 2 O): λ max> 23 (CH 2 in Bu-TOH, ref), 4.37 (t, NCH 2); 8.79 (s, Hg).

Example 20. Preparation of (S) -9- (3,4-dihydroxybutyl) guanine d-camphorsulfonate___

A mixture of (S) -9- (3,4-dihydroxybutyl) guanine and d-camphorsulfonic acid was treated as described in Example 19 to give 81% of the title compound, m.p. 170-2 ° C (95% n-PrOH + EtOAc).

1 H NMR (D 2 O): 4.38 (t, NCH 2); 8.67 (s, Hg).

30 68054

Biological tests

The herpesvirus inhibitory activity of the compounds of the invention was tested using the methods described below. The cytotoxicity of the compounds to host cell functions was also tested.

9- (3,4-dihydroxybutyl) guanine (base) is designated below VIS 707, 9- (2,4-dihydroxybutyl) guanine (base) is designated VIS 715 and 9- (3-fluoro-4-hydroxybutyl) guanine (base) is entered in VIS 912.

I. Inhibition of Viral Reproduction in Cell Cultures The herpesvirus inhibition of VIS 707, VIS 715 and VIS 912 has been measured as spot reduction according to the method below.

A. Inhibition of herpes simplex type 1 spot by VIS 707, VIS 715 and VIS 912 Spot reduction, assay for herpes simplex type 1, was performed on Vero cells (green monkey kidney cells) as described by Ejercito et al. has described in J.Gen.Virol. 2 (1968) 357. Simple layers in 5 cm petri dishes were used and after virus adsorption test compounds were added to VIS 707, VIS 715 or VIS 912 medium. The results are shown in Table 1.

table 1

Inhibitory effect of VIS 707, VIS 715 and VIS 912 on herpes simplex type 1 spot on simple Vero cell layers______________

Concentration Compound Inhibition (^ μM) (%) 1 (VIS 707) 65 5 (VIS 707)> 90 100 (VIS 715) 90 10 (VIS 912) 30 50 (VIS 912) 85 100 (VIS 912)> 90 n 31 68054 B. Spot Inhibitory Effect of VIS 707 and VIS 912 on Herpes Simplex Type 2 _______ The spot reduction test for herpes simplex type 2 was performed as in Example A. The results are shown in Table 2.

Table 2

Inhibition of a spot isolated from VIS 707 and VIS 912 herpes simplex type 2 patients by simple Veroko layers _____________

Concentration Compound Inhibition (/ UM) (%) 10 (VIS 707) 85 20 (VIS 707)> 99 100 (VIS 707) 100 100 (VIS 912) 80 II. cytotoxicity

The cytotoxicity of VIS 707, VIS 715 and VIS 912 was studied as shown in Table 3. Stenberg described the method used (Biochemical Pharmacology, Vol. 20 (1980), 1005-1008). It is seen that VIS 707 did not significantly affect cell growth at 100 μM. However, at much lower concentrations, 5 / UM herpes virus growth was inhibited by more than 90% (see Table 1).

Table 3

Cytotoxicity of VIS 707, VIS 715 and VIS 912 expressed as percentage reduction in cell growth after 48 hours of incubation _____

Concentration Compound Percentage (μM) reduction in Vero cell growth expressed as number of cells 100 (VIS 707) 10 100 (VIS 715) 29 100 (VIS 912) 30 68054 32 III. Animal Experiments A. Studies of corneal herpes in rabbits have shown that VIS 707, as an external preparation, according to Example 30, has a therapeutic effect.

The method used has been described by Alenius et al. / Acta Ophthalmologica Vol. 58 (1980) 167-173 /. Treatment was started 3 days after infection and continued for 5 days. All treatments were given four times daily. The results are shown in Table 4, where 0 means normal cornea and 3 means extensive corneal ulcer.

Table 4 Effect of 3% VIS 707 ointment on superficial corneal inflammation in rabbits___________ Days: 3 5 7 10 12

Severe corneal inflammation 3% VIS 707 latch 1.0 0.75,000

Placebo ointment 0.6 1.6 2.5 0.4 0.6 B. Experiments with herpes zoster in guinea pigs have shown that VIS 707 has a therapeutic effect in the topical preparations of Example 30. The method used has been described by Alenius et al. (Archives of Virology 58 (1978) 277-28EJ). Treatment was started 24 hours after infection and continued for 4 days. All treatments were given four times a day. The results are shown in Table 5.

Table 5

Therapeutic effect of VIS 707 on healing time and cumulative outcome ____________________

Treatment Healing time, days Cumulative result

Mean S.D. Mean S.D.

5% VIS 707- 6.6 1.8 9.0 4.1 salva

Placebo ointment 9.8 1.5 19.8 4.4

II

68054 C. Experiments using systemic HSV-2 infection in mice have shown that the oral compound VIS 707 has a therapeutic effect. The method used has been described by Olsen et al. / The Journal of Infectious Diseases Vol. 137 (1978) 428-436 /. Treatment was started one hour after infection and continued for 10 days. Mice were treated orally twice daily with 75 mg / kg VIS 707 or phosphate-buffered saline as placebo. The results are shown in Table 6. VIS 707 treatment reduced ultimate mortality and increased the mean number of days to death (MDD).

Table 6 Effect of VIS 707 treatment on mortality and mean day of death in intraperitoneally infected herpes simplex type 2__

Treatment Number of deaths /% Average number of deaths tested VIS 707 3/10 (30) 13.7

Placebo 10/10 (100) 9.3

Animal experiments showed that VIS 707 had a therapeutic effect on corneal ulcer, cutaneous herpes infections and systemic herpes infections, as shown in Tables 4, 5 and 6.

The experiment described in C was also performed on R - (+) - 9- (3,4-dihydroxybutyl) guanine, hereinafter referred to as (R) -VIS 707, as follows. A comparison was made with respect to the efficacy of placebo and the efficacy of the previously known compound acyclovir, i.e. 9- (2-hydroxyethoxymethyl) guanine.

Female NMRI mice weighing 15.0 ± 0.9 g were inoculated intraperitoneally with 10 PFU of herpes simplex type 2 virus.

They were divided into five groups of 10 mice each.

34 68054

Two groups were given (R) -VIS 707 and two groups were given acyclovir. A dose of 15 mg / kg body weight was used for both drugs.

Treatment was started one hour after inoculation and the compounds were administered orally twice daily for 10 days. The control group was given placebo only.

The mortality of the group given (R) -VIS 707 was 30%, while the mortality of the group treated with acyclovir was 70%. The results are shown in the following table.

Table 7 Effect of (R) -VIS 707 and acyclovir, at a dose level of 15 mg / kg body weight orally, and placebo effect on systemic HSV-2 infection in mice

Test compound Mortality in%, days after inoculation 7 8 9 10 11 13 14 17

Placebo 20 50 90 90 acyclovir 10 20 40 40 70 70 (R) -VIS 707 10 10 20 20 30 30

»I

35 6 8 0 5 4 In the case of uptake in the presence of guinea pigs, the presence of viruses, selective herpesviruses, etc. Sädana sjukdomar innefattar säväl vanliga infektioner soin neoplastic sjukdomar, dvs. cancer.

The effect of the virus is to reduce the effect of the intracellular end of the cellular and subcellular cells. The pathogen in the cell is a combination of viruses and viruses. Medan en del virus orsakar en allmän förstöring (dödande) av vissa Celler, kan andra transformera Celler account neoplastiskt tillstand.

Viktiga allmänna virus infection is associated with herpes dermatitis (inclusive herpes labialis), herpes keratitis, herpes genitalis, herpes zoster, herpes encephalitis, infectious mononucleosis and cytomegalovirus infection, bustling orsakas av virus som hör account herpesvirusgruppen. And the virus is infected with influenza A and B, or the influenza A and B virus.

I do not undertake any viral infection with viral hepatitis and, in particular, hepatitis B virus infection.

Effective and selective antiviral drugs are used for the treatment of endangered species, and the virus is used for the treatment of endangered species.

Otaliga olika virus av säväl DNA- som RNA-typ har pävisats orsaka cancer i djur. Effects of carcinogenic chemicals can result in activation of latent cancervirus. It is possible for a cancervirus to be co-infected with cancer.

The mannic pharyngeal pharyngeal disease, the leukemia, sarcoma, bronchial cancer, Burkitt's lymphoma, nasopharyngeal cancer and cervical cancer are indicated by RNA-cancervirus and herpesvirus as indicative. In addition, a selective inhibitor of a cancerous virus and a functional function can be used to prevent cancer.

36 68054 For 9- (4-hydroxybutyl) -guanine is described in Publication Chem. Pharm. Bull. (1 969) 1 268-1 270 och Agr.

Biol. Cheml, 37 (1973) 2037-2043. In the case of antiviral or viscous substances, the pharmacological activity is exacerbated by the action of these conditions.

U.S. Pat. No. 4,199,574 to include a group of substituents on the purifier of the formula R1 N ^ L J___N - CH-X-CH - CH-R5 D 2 ^ N ^ lii R 16 3 1 4 Rö RJ Rq color X är syre eller svavel; R is a halogen, hydroxy, alkoxy, azide, thio, alkylthio, amino, alkylamino or dialkyl-2 amine; R is, halogen, alkylthio, acylamino, amino or 3 azide; R is used, such as hydrogen or cyclic alkyl, hydroxy-4-alkyl, benzyloxyalkyl or phenyl; R is a substance, hydroxy or alkyl; R5 is selected from the group consisting of hydroxy, amino, alkyl, hydroxyalkyl, benzyloxy, benzoyloxy, benzoyloxymethyl, sulfamoyloxy, phosphate carboxypropionyloxy, raked and cyclic acyloxy with 1-8 cholatomers, e.g. acetoxy or a carbamoyl group substituted with NHCO-Z, color Z is alkyl, aryl or aralkyl optionally substituted with a sulfonyl, amino, carbamoyl or halogen; The mixture is alkyl or alkyl, which is attached to the residue X and R, R "^, R ^ and R ^ is the equivalent of R * 'is amino or methylamino, and R is hydrogen or hydroxy. The antiviral activity was a class of DNA and RNA virus, 9- (2-hydroxyethoxymethyl) guanine and 2-amino-9- (2-hydroxyethoxymethyl) adenine, which are exempt from the specific activity.

For the purposes of this Regulation, this Regulation shall apply

It 37 68054

HN I I

L I-N - CH0-CH-CH-CH „OH I

V2 color and R2, which is dirty with oil, hydrogenated fluorine, fluorine or fluorine, fused to R ^ or R ^ is used and fused to R2, R2 or R2 is fused to fluorine or R2, R2 lika, och fysiologiskt acceptabla salter eller optiska isomerer därav.

It has been shown to have hundreds of uptake and antiviral effects and to inhibit all viral functions, including tumor crystal function and viruses.

Uppfinningen ästadkommer saledes en förening och physiologiskt acceptabla salter därav, vilka föreningar är användbara vid therapeutik och / eller prophylactic behandling av virussjuk-domar och kan property användbara vid therapeutisk och / eller prophylactic behandling av cancer förorsak.

In order for an effective selective antiviral to be used as an acceptable biofinger, the selective inhibitory network must be specific to the viral function of the virus in a small number of cases.

In particular, the use of an over-the-counter treatment of a herpesvirus infection occurs. In herpesvirus can be Herpes simplex typ 1 och 2, varicella (Herpes zoster), virus with orsakar infectious mono-nucleos (dvs. Epstein-Barr virus) and cytomegalovirus. Bety-delsefulla sjukdomar orsakade av herpesvirus är herpes dermatitis (inclusive herpes labialis), herpes genitalis, herpes keratitis, herpes encephalitis and herpes zoster.

That you may be affected by the presence of cancer by cancer and tumors, in particular 68054 38 at the time of transmission of the virus. Derma effekt kan erhällas pä olika sätt, dvs. genome inhibering av transformationen av virusinfekterade Celler account tumörer och genom inhibering av virid frän transformerade Celler account andra normala Celler och genom att stoppa tillväxten hos virustransformerade Celler.

That annvation is a method for the preparation of up to-finning with the inheritance of a transformer cell using a specific herpesvirus enzyme, such as a thymidine-kinase cell.

Other possible conditions for the use of chemotherapy for the treatment of cancer and the treatment of leukemia, lymphoma of the lymphoma and Hodgkin's disease, sarcoma, cancer, nasopharyngeal cancer and cervical cancer And the other anthelmintic for the treatment of cancer with chemotherapy for the treatment of multiple myeloma and cancer in the lung (as well as bronchodilators), mage, lever, tantock, urinbläsa, läppar, ben, njurar, ä , ändtarm, spottkörtlar, mun, matstrupe, testiklar, hjärna (och skalle-ryggmärgshinnor), sköldkörtel, gailbläsa (och -gängar), näsa, larynx, bindväv, penis, vulva, vagina, liv-moderns centrala del och Tunga.

The formulation of formula I is described in A. The method of treatment of the oral or viral form of the virus in the form of an incidental mammal, varvid det sälunda infekterade djuret ges en therapeutisktektiv mängd av enförening med formuleln I eller ett physiologiskt acceptabelt sait därav, B. för inhi virus, a specific herpesvirus, and an incidental människa, genom att ge djuret som är i behov av en sädan behandling en förening med formuleln I eller ett physiologiskt acceptabelt sait därav i en mängd som är tillräcklig för inhibering av nämnda förökning,

II

39 6 80 5 4 C. for the treatment of viral inducers of neoplastic sjuk-domar i djur inclusive människa, genome inhibering av till-växten av Celler som visar Virala functioner, genom att ge sälunda infekterade djur en therapeutikitektektiv mängd av en förening med formuleln I eller with the physiologically acceptable site of the site, D. for the ingestion of viral transformers from the viral transformer Cell and the inclusive människa, genome att ge ett djur som är i behov av en sadan behandling en förening med formeln I eller et physiologiskt acceptabelt sait därav i en mängd som till som for the purpose of inhibiting the presence of the virus, E. for the treatment of the virus-inducing neoplastic suction domain and the inclusion of the mucous membrane, the genome of the use of the virus-inducing neoplastic virus, the genome and which are used for the purpose of insertion in the presence of the neoplastic suction cup and in the case of inclusions. människa, genom att ge djuret en terapeutiskt effektiv mängd av en förening med formuleln I eller dess fysiologiskt acceptable salt.

This is not the case with the use of up to a finning form.

O

--Ί - ch2-ch-ch-ch2oh 2 R, R2 color och R2, som är lika eller olika, vardera betecknar väte, hydroxi eller fluoro, förutsatt att R1 och R2 är väte da Rj och R2 är olika, och förutsatt att R 2 and R 2 are hydroxyl or fluorine, and R 2 and R 2 are hydrogen, inclusive physiologically 40 6 8 0 5 4 are acceptable salts and optical isomers.

In this case, the subgroup for which the uptake is carried out, the precursor of which is a group and R2 and the formula I are hydroxy. In this case, the subgroups of the group are selected from the group consisting of R 1 and R 2 and form I and fluorine.

In particular, it is possible to define the group R 1 and R 2 in such a way as to give a specific specific value, including the same and an optical isomer, to give up the following: 9- (3,4-dihydroxybutyl) guanine 9- (2,4 -dihydroxybutyl) guanine 9- (2,3,4-trihydroxybutyl) guanine (erythro and threo).

9- (3-fluoro-4-hydroxybutyl) guanine 9- (2-fluoro-4-hydroxybutyl) guanine 9- (2,3-difluoro-4-hydroxybutyl) guanine (erythro and threo).

For example, 9- (3,4-dihydroxybutyl) guanine is used as an enrichment.

For example, the formula I and the asymmetric risk center. Följaktligen existerar de i tvä eller fyra optiska former, och alla dessa former faller inom ramen fuprinningen.

Framställningsmetoder Föreningarna enligt uppfinningen kan framställas medelst nägon av feljande metoder A-E.

A. For the purposes of formula 41 68054 "N I I,

L J___N - CH-.CH (R1) CH (R0) COOR II

H2N ^ V N ^ 2 12 vari R1 and R2 haranan angivna betydelse, och color är väte, alkyl med 1-8 cholatomer, substituted or substituted with phenyl or benzyl, reducing.

Reductions can be carried out with borohydride, aluminum hydride, andra hydride reduction media, sodium and ethanol, with catalytic hydration or with liquid and other methods for reducing the ester to alcohol. Reductions are used for organic solvents, isopropanol for metal borohydride, dioxane for aluminum hydride or ethanol for catalytic hydration. The reaction temperature is maintained at 0 ° -50 ° C for 1 hour - 3 times.

B1. For the purposes of this Regulation

Y

”| 5

JL '_N - CH ~ CH - CH - CH-OR IV

2 I I 2 '6 H OR3 4

OR H

4 3

OR OR

H F

F H

F F

color Y is OH or a group X in which the genomic hydrolysis can be carried out on a hydroxyl group, chlorine, Bromine, iodine or a group -SH, -OR2, -SR2 or -SO2R2, i form a formulation R2 is alkyl of 1-8 cholatomer, fluorinated alkyl of 1-8 cholatomer, alkylaryl, benzyl or aryl, and R3, R4, R5 and R6 are lyc or olic and betecnar derivatives, -COR or -SO2R, color R is an angylated betydelse, hydrolyser.

42 68054

The reaction is carried out under water with a mixture of chlorine or sodium hydroxide, at a temperature of 20 ° -100 ° C under 1-24 ° C.

B2. The method B1 is defined for the formulation of formula IV, 3 4 3 5 color using the R and R columns or the R and R columns of the cyclic ketal as defined in Method E, or the cyclic carbonate ester, hydrolyser.

Reaction conditions for hydrolysis are described in Method B1.

C. In the case of formula R7 »A» Λ-ώ

I 11 R

condenser with formulation X2 - CH0 - CH - CH - CH_R10 2 r I I - 2 H R8

R9 H

R9 R8

H F

F H

F F

_J

color R7 is a silyl group having a trimethylsilyloxy or tert-butyldiphenylsilyloxy or a group Y as defined in Method 11 B1, R is an silyl group having a trimethylsilyl or a tert-butyldiphenylsilyl or a group R8 and a method R8, 9 R 10 is defined as 8, R or R is a residue or an alkyl group having a trimethylsilyloxy or a tert-butyldiphenylsilyloxy group or a group of z \ z2 or Z3 which is a residue or a halogen group containing chlorine, bromine or iodine, OR OR 4 of 5 groups definitions of method B1 or 2 43 6 8 0 5 4 method B2, and X are in the group consisting of chlorine, bromine, iodine 2 2 or group SO ~ R, color R is defined in method B1, * 7 11 varefter group R -R hydrolyser.

The condensation can be carried out in an organic solution with dimethylformamide or ethanol and at a temperature between 0 ° and 100 ° C under 1 hour and 3 times with the addition of potassium carbonate.

For example, the method B1 is used as a reaction medium for hydrolysis, for which a silyl group is used, and trimethylsilyloxylated hydrolysers are used, for example, with hydrogenated butylated sodium hydroxide. the temperature is 0 ° -50 ° C under 1-24 ° C.

D. For R1

N χ - NH “CH 2 - CH - CH - CH 2 R 2 VII

'11> i3 4 '

H H

R3 R3

F F

H il

F F

L. _ 8 9 10 11 color R, R, R, R and R are defined by Method C, 2 R -R hydrolysers.

3 ringluts in the genome of a group of myrrh or 4 derivatives of an orthoform tester, varefter group 7 11 44 6 8 0 5 4

The reaction mixture is treated with organic formates or ethanol and at a temperature between 20 ° and 210 ° C under 1 to 10 times.

E. In the case of formula R7 N * s XI1

N I I

M J-NH is a condenser with a formulation of HN ^ N ^

I i -j R

X2-CH0-CH9-CH - CH0 or X2-CH9-CH-CH9-CH9 or 2 2 | | 2 2 | 2 | 2 0 0 0 .0 \ / \ / R1 / CV2 R1 / V2 X2-CHn-CH - CH-CIUOR5 2 | | 2 0 0 \ / cv -j / \ 1 2

R 'R

1 12 in a form R is as defined in Method A, R is alkoxy 17 11 2 with 1-8 cholaters or R; R, R and X are as defined in Method C and R ~ * are as defined in Method B1, which represents a hydrolysis of the group consisting of the compounds of formula I, wherein R1 or R2 is hydroxy,

Reaction conditions for condensation are described in Method C.

In particular, methods A-E can be used to describe the enantiomeric envelope or the lower and enantiomeric isomers.

Därtill kan en oma enantiomer isomer erhällas frän enantiomera blandningar pä i och för sig kända sätt.

The materials and methods A-E are used as an alternative to the method in question.

tl 45 68054

Salter

The physiologically acceptable method of analysis is based on the genome and the method used. Salterna is not available. Metalsalter kan framställas genom att omsätta en metallhydroxid med en förening enligt uppfinningen. Exempel head metal salt can be found in lithium, sodium and potassium. That the same metal salt is used in the presence of a solid metal site. Syrasalter can be used in the genome to form an enrichment mixture with sugar, HCl, HBr, ISO4 or organic sulfonyl.

Pharmaceutical products

In the clinical administration, the administration of the active ingredient in the form of an oral formulation, orally, nasally or by genomic inhalation and in the form of a pharmaceutical preparation in the form of an active ingredient in a formulation of a pharmaceutical formulation or a pharmaceutical formulation a pharmaceutically acceptable form of food, which can be fast, cheap or free of charge, or of a capsule and of the same aspect as the uptake. Föreningen kan även användas utan bärarmaterial. Some exemptions from pharmaceutical formulations can be used in the form of tablets, droppers, powders, powders, topical applicators, powders, gels, lubricants and suspensions, aerosol dispensers for inhalation, nasal sprays, liposomers, etc.

The active substance is preferably 0.01 and 99 or 0.1 and 99% by weight, respectively. after 0.5 och 20% for oral administration for injection and between 0.1 and 50% for oral administration for oral administration.

Beredningarna är företrädesvis i form av dosenheter. The use of sterile cells in sterile form.

For the manufacture of pharmaceutical products in the form of oral dosage forms for the purpose of oral administration, the active ingredients of which may be used rapidly, 46 6 8 0 5 4 powder form, e.g. lactose, sackaros, sorbitol, mannitol, starch sasom potatisstärkelse, maisstärkelse, amylopectin, laminaria powder or pulp powder from citrus fruit, cellulose derivatives or gelatin bildning av tab-letter eller kärnor för dragöer. Om drageer önskas, kan kärnorna överdras med t.ex. concentrates of sugar, in which case the gum arabic, talc and / or titanium dioxide or alternatives to the film as a substance are added to the organic organic matter or bleached organic organic matter. Färg-ämnen kan tillsättas dessa beläggningar, t.ex. for the purposes of this Regulation, the active substance. For use in the manufacture of gelatin capsules, the gelatin capsule and t.ex. glycerol and mucus-like or liquid capsules, which may contain active substances in the form of carbohydrates or in the presence of oil, säsom t.ex. sesamolja, olivolja eller arackisolja. The gelatin capsule contains granules of the active substance in the form of a powder, lactose starch, sackose, sorbitol, mannitol, starch (i.e. potassium starch, corn starch or amylopectin), cellulose derivatives or other gelatine. lubricant.

For the purposes of this Regulation, the assets of the assets concerned shall be replaced by the same number of items. In order to adjust the weight of the active tablet, it can be used as an active ingredient in the form of granules and a mixture of granules in the form of a batch of materials. The assets of the company may be adjusted by the amount of the relevant tab of the t.ex. fats and waxes or resin tablets and tablets are made of oily material, or a physiologically inert plastic material.

For example, oral administration for oral administration may be in the form of an elixir, syrup or suspension, t.ex. a solution of about 0.1 to 20% by weight of active substance, socker and solution of ethanol, water, glycerin, propylene glycol and other aromatic substances, tl 47 68054 sackarin och / eller carboxymethicellulosa som disppergededel.

For parenteral administration, genomic injection can be carried out with the addition of active substances or physiologically acceptable salts, in a concentration of 0.05 to 10%, and preferably with stabilization and / or buffering agents. The dosage form of the solution is incorporated into the ampoule.

For topical administration, in particular for the treatment of herpesvirus infections in the head, in the body and in some cases, there is a complete formulation in the form of an ointment, ointment, gel, suspension, cream or liquid. Game with active assets can be varied, t.ex. with 0.05 to 20% by weight of active substance. It is intended for the administration of the above-mentioned genome to the active substance, including isopropanol, glycerin, paraffin, stearyl alcohol, polyethylene glycol, etc. The pharmaceutical acceptor has the right to use a chemical absorption promoter. Exempel head absorptionspromotorer är t.ex. dimethylacetamide (U.S. Pat. No. 3,472,931), trichloroethanol or trifluoroethanol (U.S. Pat. No. 3,891,757), best alcohol and bleaches (British Patent No. 1,001,949).

The dosage and the active assets of the administrators can be varied in such a way that the interval between and above the factor factor is t.ex. infections, patients, etc. och den möjligen justeras individually. In this case, the interval between the first or second instance of the administration is about 0.1 mg to 2000 mg or about 1 to 2,000 mg or more from 1 to 2,000 mg for the top administration, 50 to 2,000 mg to 100 to 1,000 mg. mg for oral administration and 10 to 2000 mg or 50 to 500 mg for injection.

I allvarliga fall can be found in the case of fem- till thiophaldehyde. In all cases, 500 to 1000 mg of the active ingredient are added.

48 6 8 0 5 4

The pharmaceutical composition of the active ingredient may be formulated in such a way that the dosage is at intervals between the dosage and the multi-dose enhancer.

The physiologically acceptable metabolite of the herpesvirus may be affected by the use of the herpesvirus. Formulations of the formulation and physiologically acceptable methods of treatment and therapeutic or prophylactic treatment of viral infections.

A particular aspect of uptake is the use of a formulation according to formula I or a physiologically acceptable site for the treatment of herpesvirus infections.

Exemption from the application of the exemption is illustrated by the provisions of this Regulation.

I. Framställning av utgängsmaterial

Example 1. Framställning av 4-Bromo-2-hydroxybutylsyraetylester

Br-CH 2 CH-, CH-COOCHnCH-, λ 2 i 2 ό

OH

2-hydroxybutyrolactone, British Patent 688 253 _ / C. A. 48, pp. 3996 (1 954), (5.1 g) was added to 10 ml of ethanol and the solution was stirred with hydrogen bromide at 0 [deg.] C. After that, the fat was heated at room temperature for a period of 66 minutes. The addition of a mixture of reducing agents and a mixture of ether-derived extracts with the addition of sodium derivatives Sodium sulphate After distillation at the distillation stage, at a temperature of 12 mm Hg, the fraction of the sorghum boils at 109-112 ° C with a weight of 3.79 g, which is obtained in 2 copies.

il «68054

Exempel 2. Framställning av 4- (2-amino-6-chloropurin-9-yl) -___- 2-hydroxybutyrsyraethyl ester _

Cl

"l-N

L 1 11

H2K I (j) H

CH CH 2 CH-COOCH 2 CH 3 2-Amino-6-chloropurine (0.509 g, 3.00 mmol), 4-Bromo-2-hydroxybutyr cyrethyl ester (0.633 g, 3.00 mmol; freely exempted from 1) and aqueous potassium carbonate (0.415 g) , 3.00 mmol) are stirred in 10 ml of dimethylformamide and bleached at room temperature under 65 g. The filtrate is filtered and induced to a temperature of 0.1 mm Hg. The crystals of the residue are triturated with 8 ml of chloroform and the material is filtered off with 2 ml of chloroform. The material is triturated with 5 ml of water and the material is filtered with 2 ml of water. Crystallization from 11 ml of ethanol gave 0.360 g of product. Smp. 16 3-1 6 4 ° C (ej corrugated). UV spectrum (ethanol): λ Inax / nm) 311, 248. Analyzes: Fnetnet: C 43.90; H 4.78; Cl 11.72; N 23.52; O 15.90%. Beräknat föreningen C. .H. .ClNr-O 2: C 44.08; H 4.71; II I h O 3

Cl 11.83; N 23.37; 0 16.01%.

Example 3. Preparation of 4- (2-amino-1,6-dihydro-6-oxopurin-__-9-yl) -2-hydroxybutyrsy

Jk HN. li

JL 1 II

u

h, N, OH

ί I i

CH2CH2CH-COOH

4- (2-Amino-6-chloropurin-9-yl) -2-hydroxybutyrsyrethyl ester (1.40 g, 4.67 mmol; preferably 2-fold) is added to 100 mL of 1M water to reflux the salt under 2.5 g. The solution is obtained after a dip of about 10 mm Hg. Watermelon (25 ml) was added at 6 80 5 4 times and the starting material was removed. This procedure was performed on the ginger. The residue is triturated with 150 ml of acetone and half of the material filtered off, with a yield of 1.38 g. The resulting product (1.27 g) was taken up in 5 ml of water, filtered through and treated with 2.5 ml of water. The pH of the filtrate is adjusted to 6 to 7 times with fast sodium bicarbonate. The aqueous solution is filtered and the material is taken up in 7.5 ml of water. 0.4 ml of acetic acid is filtered off. After cooling to 0 ° C, filtering is carried out with 3 ml of water. Crystallization from 75 ml of water gave 0.68 g of product. Smp. 200 ° C (sonication). UV spectrum (0.1M salts): (nm) 279, 254; UV spectrum (0.1M sodium hydroxide); (nm)

IllclX ulciX

269 (infl.) Analysis for C18: C 42.63; H 4.41; N 27.74; O 25.30%. Found for C 9 H 18 N 2 O 2: C, 42.69; H 4.38; N 27.66; O 25.27.

Example 4. Preparation of 4- (2-amino-1,6-dihydro-6-oxypurine-9-yl) -2-hydroxybutyrylethyl ester

O

-OF

L X j CH2CH2CpHCOOC2H5

OH

4- (2-Amino-1,6-dihydro-6-oxypurin-9-yl) -2-hydroxybutyrsra (2.00 g, 7.9 mmol; free from 3) was added with 500 mL of ethanol. Blandningen mättades med klorvätegas, först utan avkylning och därefter med avylylning i is-vatten. The total time is about 15 minutes. Blandningen uppvärmdes därefter längsamt account rumstemperatur och fick stä över natten. After the addition of the solution is carried out with a solution of 25 ml of ethanol per gang, the color of the preparation is then reduced to 25 minutes. The mixture is then added to 12 ml of water and the pH is adjusted to 6-7 with the addition of sodium bicarbonate. Fälningen avfiltrerades, teaspoon 51 68054 with 2 ml of water and piercing under vacuum, coloring 1.60 g. Crystallization from ethanol yields the product, m.p. 161-163 ° C (that the analysis is at 162-162 ° C).

Analysis: C: 46.96; H 5.35; N 24.77; O 22.60%. Found for C 18 H 19 N 2 O 2: C, 46.97; H 5.38; N 24.90; O 22.74%.

Example 5. Preparation of 4- (2-amino-6-chloropurin-9-yl) methyl -_-1,3-dioxane _____

Cl

^ ^ -N

H2 ^ / CH_ — CH

CH X0 - CH2 ^

Equimolar compounds include 2-amino-6-chloropurine, 4-bromomethyl-1,3-dioxane (framställt enligt Price C.C. J. Amer. Chem. Soc. 1950, 72, 5335-6) and wattenfrit potassium carbonate blandades i torr dimethylformamide.

After 7 days of blending at room temperature, the filtrate is bleached and the filtrate is removed under reduced pressure. The residue is triturated with ethanol and the other material is filtered off. The filtrate was removed to give a crystal and crystallized fast to give a crystalline mixture of chloroform,

colors färglösa stavformade kristaller erhölls. Smp. 169-70 ° C

(ej corrected) UV spectrum (salts 0.01 mol / l): λ (nm) 308, 245. UV spectrum (ethanol): λ (nm) 310, 247. M.S: 11.2

ITlclX

a J., m / e (int.): 271/269 (0.19 / 0.55), 226/224 (0.11 / 0.28) 212/210 (0.33 / 0.82) 171 / 169 (0.35 / 1.0).

Example 6. Preparation of 2-fluorobutyrolactone 2-Bromobutyrolactone (15 g) and silver fluoride (23 g) were bleached under 24 times with acetonitrile (150 ml).

After cooling to room temperature, the filtration mixture is stirred and treated under vacuum. The mixture was taken up in ethyl acetate (200 ml) and water (6 x 50 ml). The aqueous phase is taken up in ethyl acetate (80 ml) and combined with ethyl acetate in the form of magnesium 52,68054 sulphate and starting, 2-fluorobutyrolactone (21%). 1 H-NMR (acetone-d 6, Me 4 Si) δ: 5.02 and 5.95 for CHF (triplet, J = 8 Hz. 0 ^ ^ = 51 Hz).

Example 7. Fractionation of 4-bromo-2-fluorosubstituted ethyl ester The bromide was stirred under ca. 30 minutes to give 2-fluorobutyrolactone (1.8 g) in ethanol (15 mL) at 0 ° C. Reaction-bleaching at room temperature for 3 hours, under reduced pressure. The mixture is stirred in water, neutralized with sodium carbonate and extracted with ether. Ethylation was carried out with the addition of sodium sulfate, addition of sodium sulfate and induction, coloring of 4-bromo-2-fluorosubstituted ethyl ester (1.26 g, 34%). n ^ ° = 1.4698.

1 H-NMR (COCl 3, MeSi) δ: 1.3 (t, J = 7Hz, CH 3); 2.1-2.8 (m, CH 2); 3.55 (t, J = 7Hz, CH 2 Br); 4.3 (q, J = 7 Hz, CH 2); 4.75 and 5.55 (tva tripletter, J = 6Hz, CHF, JcHp = 50 Hz).

Example 8. Framställning av 4- (2-amino-6-chloropurin-9-yl) -2-fluoropyrethyl ether

Cl

, / r V

CH_CHoCH-C0oCHoCH,

i Z j / / J

F

4-Bromo-2-fluorosubstituted ethyl ester (0.23 g), 2-amino-6-chloropurine (0.19 g) and hydrogenated potassium carbonate (0.15 g) are added dropwise to dimethylformamide (3.7 ml) at room temperature under 2 parts. . The solution was filtered and induced under vacuum. The mixture was triturated with chloroform (4 + 2 mL) and treated with chloroform to give the product as a solid (0.24 g, 72%). TLC (silica gel, i-propanol-watten conc. Ammonia 8-1-1): Rf = 0f76. NMR (CDCl 3, Me 4 Si) δ: 1.26 (t, CH 2); 2.4-2.5 (m, CH 2); 4.2-4.3 (t och q, NCH 2 and CO 2 CH 2); 4.77 and 5.03 (CHF, J = 51Hz); 5.2 (NH2); 7.77 (8-CH).

ti 53 68054

Example 9. Preparation of 4- (2-amino-6-chloropurin-9-yl) -2-fluorobutanol______

Cl i XI> N / N 'H | ch2ch2chch2oh

F

Sodium borohydride (15 mg) was taken up in diethylene glycol diethyl ether (0.9 ml) and treated. The title compound was treated with lithium bromide (34 mg), followed by 4- (2-amino-6-chloropurin-9-yl) -2-fluororesuberethyl ester (100 mg). The reaction mixture is immersed at 100 ° C under 3 timetables, leaving the diphther in cross-section (10 g) with a thick mixture of salts (0.5 ml) and acid. The pH is adjusted to 6.5 with sodium bicarbonate, and the solution is taken up in vacuo and treated with methanol-chloroform (40-60, 10 ml) and filtered. The solution was chromatographed on a silica gel column eluting with a methanol-chloroform gradient to give the desired product as an oil (52 mg, 60%) by TLC (silica gel, methanol-chloroform 40-60): R = 0.66. UV (ethanol) λ: 310 and 247 nm.

ς m cix

Exempel 1 0. (s) -4- (2-Amino-6-benzyloxy-piper-9-yl) -1,2-O-isopropyl-1,2-dihydroxybutane _______ O-CHoCcHc T 2 6 5 XX> ΗΛΓ N i1 x CH - CH - CH - CH0 (S) 2 2 | | 2 ° x / °

C

/ \ CH3 CH3

__ - TT

54 6 8 054 (S) -1,2-O-Isopropylidenebutane-1,2,4-triol (framställd ur (S) - (-) - maleic acid en Hayt Hayi H. et al. J. Amer. Chem. Soc. 1973, 92, 8749-8757) mesylated (by the method of Crossland RK ooh Servis KL Org. Chem. 1970, 35, 3195) ooh (S) -4-O-methanesulfonyl-1,2-O-isopropylidenebutan-1, 2,4-triol was used without further purification.

2-Amino-6-benzyloxypurine (Bowles WA et al. J. Med. Chem. 1963, 6, 471-480) and tetrabutylammonium hydroxide are refluxed with chloroform and extracts (MgSO 4) and induced to give 2-amino 6-benzyloxipurintetrabutylanunoniumsalt.

2-Amino-6-benzyloxypurine tetrabutylammonium salt (300 mg, 0.62 mmol), (S) -4-O-methanesulfonyl-1,2-O-isopropylidenebutane-1,2,4-triol (140 mg, 0.62 mmol) and tetrabutylphenium iodide (22 mg, 0.06 mmol) in chloroform (10 mL) were refluxed under 24 tincture under vacuum. Chromatography on silica gel (silica gel, methanol-chloroform 5-95 eluent) gave 72 mg (31% yield) of the title compound.

After washing with water, the mixture was evacuated and treated with ether to give a solid which was first purified by TLC (silica gel, CHCl 3 -MeOH 20-1, Rf 0.33). 1 H NMR (CDCl 3): 1.36 and 1.44 (2s, 2x3 H) isopropylidene group; 2.1 (m, 2 H) 2 H 2; 3.6-4.4 (m, 5H) 2H1, 1H3, 2H4; 5.15 (bred s, 2H) NH 2; 5.64 (s, 2H) benzyl CH2; 7.5 (m, 5H) phenyl; 7.73 (s, 1 H) Hg.

II. Preparation of 9- (3,4-dihydroxybutyl) guan

O

L l_nI

h2n ^ n / CHo-CHo-CH-CH2H (VIS 707)

£. £. | Z

OH

To a suspension of ethyl 4- (2-amino-1,6-dihydro-6-oxopurin-9-yl) -2-hydroxybutyrate (preferably 4) isopropanol 55,68054 was added to sodium borohydride and reflux solution. -rades över natten (minst 8 tinunar). Saltsyra tillsattes tills en klar lösning erhölls (neutral pH). After cooling to the end of the day, the water is heated at a minimum temperature of 0 ° C. The first filter is used. The filtrate was triturated under reduced pressure and treated with salts (1 mol / l) and adsorbed onto a cationic resin (Dowex 50 W, H + form). The resin is treated with water and eluted with 5% ammonium hydroxide. Elueringsmedlet indunstades, varvid ett kristallint fast ämne erhölls, som kristallerades ur vatten och gav färglösa hunger. Smp. 260-1 ° C (sonication) (corrected) UV spectrum (salt 0.01 mol / l): λ (nm) 277, 253 (ε = 11500) MS: 11.2 a J. (int.): 239 (M +, 0.13), 222 (0.19), 221 (0.11), 152 (0.43), 151 (0.56), 44 (1.0).

Exempel 12. Framställning av 9- (2,4-dihydroxybutyl) guanine HJi I I (VIS 715)

CH 2 CH-CH2CH2OH

OH

4- (2-Amino-6-chloropurin-9-yl) methyl-1,3-dioxane (preferably 5) is added to the salt (1 mol / l) and refluxed under 4 times. The alkaline solution is treated with ammonium hydroxide and induced by reduction under reduced pressure. The reaction was carried out in water and then with preparative HPLC on a flash column (Bondapack Clg), eluting with bleaching with methanol and water (1: 3). These products can be used to crystallize fast. Smp. 226-8 ° C (crystallization) (corrected) UV spectrum (salt content 0.01 mol / l): λ__ "(nm) 277, 254 (ε = 10700).

Exempel 13. Framställning av 9- (3-fluoro-4-hydroxybutyl) guanin 56 68054

Hf I S (VIS 912)

N N 'F

H I I

Ch2ch2chch2oh 4- (2-Amino-6-chloropurin-9-yl) -2-fluorobutanol (52 mg) was added dropwise to refluxing 1M aqueous HCl (4.5 mL) under 50 min. The solution was stirred under vacuum, and the solution was added to water and the solution was neutralized with 1M ammonia and lyophilized. Intermediate renates with an anion exchange column (Dowex 1x2, OH).

The starting product was obtained as an own product (37 mg, 77%). NMR (DMSO-d 6) δ: 1.95-2.15 (m, CH 2); 3.3-3.5 (m, C.H 2 O); 4.05 (t, NCH 2); 4.3 and 4.5 (CHF, J = 49 Hz); 6.45 (NH2); 7.7 (8-CH); 10.6 (NH).

Exempel 14. Framställning av 9- (DL-erythro-2,3,4-trihydroxybutyl) -_guanine___ fl

HN ^ X / 'NX

I I | (VIS 925)

L Π__N-CH ~ -CH-CH-CH-OH

2 I I 2

H2N OH OH

6-Chloroguanine (0.75 g) and sodium hydride (60% in oil, 0.18 g) were added to dimethylformamide at room temperature under 1 hour. DL-erythro-2,2-dimethyl-4-p-toluenesulfonyloxymethyl-5-hydroxymethyl-1,3-dioxolane / 0.70 g, freely obtained from A. Hol, Coll. Czech. Chem. Commun Voi 44, siderna 593-61 2 (1 979 | _7 volumes and reaction bleaching at 100 ° C under 17 timetables, the genome is filtered under pressure and the reaction medium is stirred under vacuum. Refluxing with acetic acid (25 ml)). Under 2 timetables, the reaction medium is diluted and diluted with water (3 x 5 ml) and then poured into a volume of 1N salt and column Dowex 50x8 column (H +, 70 ml), eluting with water (1 liter). - to 57 6 8 0 5 4 ammonia fractionation (up to 1/10), eluting with a UV detector, heating the fraction under vacuum and using an intermediate mixture with water, up to a maximum of 230 ml of water (230 ml). ) and filtrates, coloring of 9- (DL-erythro-2,3,4-trihydroxybutyl) guanine (0.20 g).

NMR (DMSO-d 6) δ; 7.8 (s, 8C-H); 6.4 (NH2); 4.22 (N-CH2).

Example 15, Framställning av (S) -9- (3,4-dihydroxybutyl) guanine O

οΑΛ »'CH2CH2C ^ H-CH2OH (S)

OH

Preparation of (S) -4- (2-amino-6-benzyloxypurin-9-yl) -1,2-O-isopropylidene-1,2-hydroxybutane with salt syrup (1 mol / l) at 100 ° C under 1 Timme gav of (S) -9- (3,4-dihydroxybutyl) guanine.

TLC (silica gel, isopropanol-water-concentrated ammonia 7: 2: 1,

Rf 0.60), HPLC on a Bondapak Phenyl / porasil B column, methanol-water 30-70), and UV (0.1 mol / l salt salt) can be identified as a racemic reaction (VIS 707).

Example 16. Framställning av R - (+) - 9- (3,4-dihydroxybutyl) guanine Steg a: Framställning av R - (+) - 1,2,4-butanthriol R - (+) - dimethylmalate (1,62 g, 10 mmol), framställt enligt Boger, DL and Panek, J.S. J. Org. Chem. 1981, 46, 1208-10, poured into THF (10 mL) and added dropwise a suspension of lithium aluminum hydride (0.63 g, 16.5 mmol) in THF (15 mL). The reaction mixture was stirred at 55 ° C. After successful addition to water (0.62 ml), 10% sodium hydroxide (1.20 ml) and water (1.90 ml) were filtered through blends and the mixture was boiled with THF (2 x 20 ml). and filters. The filtrate was combined and reduced to a reduced column (13 Pa, 30 ° C), m.p. 6,804 followed by 1,2,4-butanthriol hydrochloride (0.7 g, 6.6 mmol), 66%.

Step b: Preparation of R - (+) - isopropylidenebutane-1,2,4-triol R - (+) - butanthriol (0.7 g, 6.6 nunol) is obtained in the form of a mixture of 1.5 g. Acetone in acetone (50 ml) was added to 3 portions of concentrated perchloride, and the sodium bicarbonate solution was added to the water (2 ml) at room temperature and then stirred for less than 10 minutes. The filtrate was filtered and filtered under reduced pressure (2.7 Kpa, 30 ° C). The reaction mixture was taken up in ethyl acetate, with the addition of sodium bicarbonate (5 ml) and brine (5 ml) and triturated over magnesium sulfate. The starting material was isolated and distilled, and the title compound was obtained as an oil (0.3 g, 2.05 mmol, 31%): m.p. 104-6 ° C at about 2.7 kPa, n ° = 1.4390.

Step c: Preparation of R - (+) - 4-bromosopropylidenebutane-1,2-diol R - (+) - isopropylidene-butane-1,2,4-triol (0.3 g, 2.05 mmol) and triphenylphosphine (0.63 g, 2.4 mmol) was taken up in methylene chloride (5 mL) and cooled to 0 ° C. N-bromosuccinimide (0.38 g, 2.16 mmol) was added portionwise at 0 ° C. After stirring at 0 ° C, hexane (15 ml) was added and then filtered through a pad of hexane (2 x 5 ml). The combined hexanes were combined with a silica gel column (5 g). Elution with hexane (15 mL) was followed by induction and distillation to give a clear oil (0.2 g, 0.96 mmol, 47%): m.p. 74-6 vid 2.7 kPa (20 mm Hg), n ^ ° = 1.4630. [.Alpha.] D @ 20 = + 27.7 (c = 20, CHCl3).

Step d: Preparation of R - (+) - 4- (2-amino-6-chloropurin-9-yl) -isopropylidene-butane-1,2-diol 2-amino-6-chloropurine (162 mg, 0, 96 mmol), R - (+) - 4-bromosopropylidene-butanediol (200 mg, 0.96 mmol) and potassium carbonate (132 mg) in DMF (10 mL). After 16 minutes of reaction, the reaction of the genome with celite and the solution is reduced to a reduced temperature of 59 68054 cerat / L (2 x Hg), 50 ° C7 · The reaction is triturated with chloroform (5 ml) and dried. olösliga substansen avfiltrerades. In addition, the color is rapidly crystallized at a fast crystalline rate, which is preferably separated from the 9- and 7-isomers. These are separated by silica gel chromatography. Elution with chloroform / methanol (15: 1) gave the title compound (106 mg, 0.36 mmol, 37%): m.p. 129-30 ° C / 07 ^ 0 = + 57.5 (c = 6'97 'chCl3).

Step e: Framställning av R - (+) - 9- (3,4-dihydroxybutyl) guanine R - (+) - 4- (2-amino-6-chloropurin-9-yl) isopropylidene-butane-1,2- Diol (100 mg, 0.33 mmol) was added dropwise to the chloroform solution (1 mol / L) and refluxed under 1 timetable. The solution was concentrated in vacuo and the reaction mixture was washed with water (5 mL) and the alkaline genome was added with water and ammonium hydroxide. After starting the crystals, the solution was added to water, giving the title compound (40 mg, 0.17 mmol, 51%). UV spectrum (0.01 mol / l chloroform): 1__ (nm) 277, 253.

^ 1 luaX

/ 07 ^ = + 30.8 (c = 0.25, watts). PMR (D 2 O ref. T. BuOH, oCH = 1.23 ppm), 1.75-2.15 ppm (m, 2H), 3.40-3.70 ppm (m, 3H), 4.15 ppm (AB, 2H) and 7.77 ppm (s, 1H).

9

Example 17. Preparation of R - (+) - 9- (3,4-dihydroxybutyl) guanine hydrochloride (R) -9- (3,4-dihydroxybutyl) guan (5.02 g, 21.0 mmol) was added dropwise. 5 M chloroform (5.6 mL, 28 mmol) is added. After crystallization at 0 ° C by lyophilization, the hydrochlorides are concentrated and quantitatively determined, m.p. ca 160 ° C (after vacuum vacuum 110 ° C).

1 H NMR (D 2 O): 61.9-2.3 (m, 2H) NCH 2 CH 2; 3.5-3.9 (m, 3 H) CHCH 2 O; 4.39 (t, 2H) NCH 2; 4.9 (s) HDO; 8.91 (s, 1H9 Hg <.).

6o 68 0 5 4

Example 18. Framställning av 9- (3,4-dihydroxybutyl) guanine

Step a: Preparation of 4- (2-amino-5-nitro-6-oxypyrimidin-4-yl) -1,2-O-isopropylidene-1,2-butanediolamine

O

i I

N NHCHnCHnCH - CH ~ 2 2 | | 2

O O

X

H 3 C CH 3 2-amino-4-chloro-5-nitro-6-pyrimidinol (3.80 g, 20 mmol), potassium bicarbonate (2.00 g, 20 mmol) and 1-amino-3,4-O- Isopropylidene dihydroxybutane (2.92 g, 20 mmol) was added to 40 mL of dimethylformamide. The solution was stirred and triturated with 50 ml of ethanol. The mixture is filtered, filtered and crystallized with 1 liter of ethanol. The essential substance is 2.29 g. 1.88 g of a crystalline solid are added to the ethanol solvent.

Step b: Preparation of 9- (3,4-dihydroxybutyl) guanine 4- (2-amino-5-nitro-6-oxo-pyrimidin-4-yl) -1,2-O-isopropylidene-1,2-butanediolamine (283 mg, 1 mmol) in 100 mL of ethanol hydrated with 5% palladium tritol and tricc at 4 atm under 72 timmar. The solution was stirred under stirring with diethoxymethyl acetate (5 mL, 31 mmol). Blandningen fick stä över natten. Add a mixture of diethoxymethyl acetate and trifluoroacetate (5 mL, 65 mmol). Blandningen fick stä i ett kyl-skäp under 5 dygn. The products were analyzed by HPLC with 30% methanol in water on a C18 column and the products were identical to those used in Method A (exemptions 11). HPLC yielded 88%.

Il

Example 19. Framställning av (R) -9- (3,4-dihydroxybutyl) guanine-d-camphorsulfonate___ si 6 8054

The mixture of (R) -9- (3,4-dihydroxybutyl) guanine (0.50 g, 2.09 mmol) and d-camphorsulfonyl (0.50 g, 2.14 mmol) was added under stirring with 1.5 g. ml of 2-propanol, saturated and purified over the course of crystallization and filtration, colored to 0. 896 g (80%) of the title compound. Crystallization from 95% to 1% propanol and ethyl acetate. melting point 1 90-2 ° C.

1 H NMR (D 2 O): δ 1.23 (CH 3 i t-BuOH, ref), 4.37 (t, NCH 2); 8.79 (s, Hg).

Example 20. Framställning av (S) -9- (3,4-dihydroxybutyl) guanine-d-camphorsulfonate_______

The purification of (S) -9- (3,4-dihydroxybutyl) guan and d-camphorsulfonyl compounds is carried out according to exemptions 19, with a yield of 81% of the title compound, m.p. 170-2 ° C (95% n-PrOH + EtOAc).

1 H NMR (D 2 O): 4.38 (t, NCH 2); 8.67 (s, Hg).

Biological test

The effect of the present invention is to determine whether the herpes virus has been tested by means of a non-essential method. Föreningarnas cellulära toxicitet pä värdcellens funktioner testades även.

Forms 9- (3,4-dihydroxybutyl) guanine (bas) are not present in VIS 707, for 9- (2,4-dihydroxybutyl) guanine (bas) is VIS 715 and for 9- (3-fluoro-4-hydroxybutyl) guanine (bas) free VIS 912.

1. Inhibition of viral infections in cell culture Föreningarnas VIS 707, VIS 715 and VIS 912 Inhibition of herpesvirus may be carried out using the following methods:

62 6 8 0 5 4 A. Föreningarnas VIS 707, VIS 715 and VIS 912 inhibering av herpes slmplextyp 1 plack_

Plackreduktionsförsöket för herpes simplextyp 1 utfördes pä Vero-Celler (Green Monkey Kidney) sasom beskrivits av Ejercito et al. and publications J.Gen.Virol. 2 (1968) 357. Monoskikt i 5 cm petriskälar användes och efter adsorption av virus tillsattes testföreningarna VISna 707, VIS 715 ellis VIS 912 till mediet. The results are shown in Table 1.

Table 1 Föreningarnas VIS 707, VIS 715 and VIS 912 inherent effect on herpes simplextyp 1 plack of Vero-cell monoskikt_

Concentration for Inhibering (/ UM) __ (%) _ 1 (VIS 707) 65 5 (VIS 707)> 90 100 (VIS 715) 90 10 (VIS 912) 30 50 (VIS 912) 85 100 (VIS 912)> 90 B. Effective effect of VIS 707 and VIS 912 on herpes simple type 2 plaque________

Plackreduktionsförsöket för herpes simplextyp 2 utfördes pä samma sätt som i exempel A. The results are shown in Table 2.

Tabell 2 Föreningarnas VIS 707 och VIS 912 inhibering av herpes simplex- typ 2 plack isolate frän patient pä Vero-cell monoskikt_

Concentration for Inhibering (/ UM) (%) 10 (VIS 707) 85 20 (VIS 707)> 99 100 (VIS 707) 100 100 (VIS 912) 80 1! 68054 63 II. Cellular toxicity For the VIS 707, VIS 715 and VIS 912 cellular toxicity tests, all of which are shown in Table 3. Methods are described in Stenberg / Biochemical Pharmacology, Vol. 20 (1980), 1005-10087. Märkbart head cell culture volume at 100 μM. At a concentration of 5; UM inhibited the presence of herpes virus infections at / 90% (see Table 1).

Table 3 Formed VIS 707, VIS 715 and VIS 912 cellulite toxicity in the case of percentage reduction in cellulose after 48 incubations__

Concentration For the purpose of reducing the amount of tax in the cell-cell field and cellular 100 (VIS 707) 10 100 (VIS 715) 29 100 (VIS 912) 30 III. Djurexperiment A. The experiment with herpes keratitis can be carried out for VIS 707 and the topical effects are 30 or less therapeutically effective. The methods described in Alenius et al / Acta Ophthalmologica Col. 58 (1980) 167-173 /. Behandlingen päbörjades 3 dagar efter injectionen och fort-sattes i 5 dagar. The aid is obtained. The results are shown in Table 4, vari 0 betecknar normal hornhinnan och 3 betecknar en utsträckt särbildning i hornhinnan.

Table 4

Effects of 3% up to VIS 707-ointment on inflammation in the horn- price hos kaniner___

Amount: 3 5 7 10 12 _Häftighet av inflammation i hornhinna 3%: ig VIS 707-salva 1,0 0,75 0 0 0

Placebo salva 0,6 1,6 2,5 0,4 0,6 64 68054 B. Experiments with herpes may be carried out for VIS 707 and topical treatments with 30 therapeutic effects. Methods are described in Alenius et al / Archives of Virology 58 (1978) 277-2887 * Behandlingen pabörjades 24 timmar efter infectionen och fort-sattes i 4 dagar. The aid is obtained. The results are shown in Table 5.

Table 5 Effects of VIS 707 on therapeutic effects and cumulative results_

Behandling Läkningstid Dagar Cumulative result __Medelvärde_S. D._Medelvärde_S «D._ 5% to VIS 707- salva 6.6 1/8 9f0 4.1

Placebo-ointment 9.8 1.5 19.8 4.4 C. An experiment with the development of a systemic HSV-2 infection in a study of VIS 707 orally administered a therapeutic effect. Methods described in Olsen et al / The Journal of Infectious Diseases Vol 137 (1978) 428-4367 * Behandlingen pabörjades 1 timme efter infektionen och fortsates i 10 dagar. Each oral dose of 75 mg / kg VIS 707 or phosphate buffered saline is placebo. The results are shown in Table 6. The use of VIS 707 reduced the mortality rate and the value of the data for the same period (MDD).

Table 6

Effects on the use of VIS 707 in mortality and in the administration of moss infections i.p. med herpes simplextyp 2_

Behandling Antal döda /% Medeldödsdagen

Antal testade VIS 707 3/10 (30) 13.7

Placebo 10/10 (100) 9.3

Djurexperiments have been performed for the treatment of VIS 707 with therapeutic effects in herpes keratitis, in particular herpes infections and 65 6580 in the case of herpes infections in Table 4, 5 and 6.

The test is carried out under point C to give R - (+) - 9- (3,4-dihydroxybutyl) guanine without the addition of (R) -VIS 707. The compounds of placebo and the effects of acyclovir, dvs. 9- (2-hydroxyethoxymethyl) guanine, are also effective.

NMR 1 in the presence of 15.0 ± 0.9 g of inoculations i.p. med 4 10 PFU herpes simplextyp 2 virus. The group is in the group with 10 mosses in the group.

Tvä grupper erhöll (R) -VIS 707 och tvi grupper erhöll acyklovir.

A dose of 15 mg / kg of crude oil is added to the package.

Behandlingen pabörjades en timme efter inoculating och för-eningarna gavs oralt tvä ganger dagligen under 10 dagar. The control group is a placebo.

Mortality in the group with erhöll (R) -VIS 707 var 30% Medan mortality in the group with acyclovir var 70%. The result is a table.

Table 7

Effects of (R) -VIS 707 and acyclovir at a dose of 15 mg / kg on oral administration and placebo on systemic HSV-2 infection and ____

Test mortality in%, after inoculation 7 8 9 10 11 13 14 17

Placebo 20 50 90 90 acyclovir 10 20 40 40 70 70 (R) -VIS 707 10 10 20 20 30 30

Claims (2)

A process for the preparation of a therapeutically active compound of the formula 0 II N HN k II 1 H n / n ^ N ^ N - CH 2 -CH (R 1) CH (R 2) CH 2 OH I wherein both R 1 and R 2 are the same or hydrogen, hydroxy or fluorine, provided that R1 or R2 is hydrogen when R1 and R2 are different and provided that R1 and R2 are hydroxy or fluorine when R1 and R2 are the same or a physiologically acceptable salt or optical salt thereof. for the preparation of an isomer, characterized in that A. a compound of the formula O 1 il 1 is reduced, - N - CHOCH (R 1) CH (R 0) COOR 1 II H 2 N 1 212 wherein R 1 and R 2 are as defined above and wherein R 1 is hydrogen, alkyl of 1 to 8 carbon atoms, substituted or unsubstituted phenyl or benzyl, B1. hydrolyzing a compound of formula Y t "| Ί 5 --N - CH.CH - CH - CH" 0R3 X '_21 | o ~ i 2 «NH OR3! R6 OR4 H j IV I OR4 OR3 j HFIFH: ff J wherein Y is OH or a group X which can be converted by hydrolysis to 67 68054 hydroxy groups such as chlorine, bromine, iodine or the group -SH, 2 2 2 2 -OR, -SR or -SOjR, wherein R is a group containing 1 to 8 carbon atoms alkyl, fluorinated alkyl having 1 to 8 carbon atoms, alkylaryl such as benzyl or aryl, and R 3, R 4, R 8 and R 6 are the same or different and represent hydrogen, -COR or, where R is as defined above Hydrolyzing a compound of formula IV as defined in method B1, wherein R and R together or R and R together or R and R together form a cyclic ketal as defined in method E, or a cyclic carbonate ester, C. condensing a compound of formula R7 J. N, N -! I jH VIIT HN- '------ N i11 with a compound of formula X2 - 0Ho - CH - CH - CH-R10 2. II 8l 2 HR R9 H R9 R8 HFFH _ F F_ 7 wherein R is a silyloxy group such as trimethylsilyloxy or tert. -butyldiphenylsilyloxy or a group Y as defined in Method B1, R is a silyl group such as trimethylsilyl or a tert-butyldiphenylsilyl or a group R6 as defined in Method B1, and R8, R9 and R19 are the same or different and mean silyl groups such as trimethylsilyloxy or tert-butyldiphenylsilyloxy or groups 12 3 Z, z and Z which are identical or different and denote halogen, such as chlorine, bromine or iodine, or groups OR 3, OR 4 or OR 8 as defined in method B1 or go to - 2 68054 in process B2, and X is a leaving group such as chlorine, bromine, 2 2 iodine or a group SO_R, in which R is as defined in process B1, 7 11, after which the groups R -R are hydrolysed, D. a compound of formula n R 'i N' <:; , NH2 i L ^ N x 1 f) HN 'NH - CH0 - CH - CH - CHO R U I i 1 2. J l q 2 R HR8 9 R H R9 R8 R R VII H F F H! FF i _ _ 7 8 9 10 11 wherein R, R, R, R and R are as defined in Method C by reacting it with formic acid or a derivative thereof such as orthoformate ester, after which the groups 7 1 1 R -R are hydrolyzed, or E. is condensed a compound of formula R7? II XII HNNH i "with a compound of formula X2-CH0-CH- ~ CH - CH0 or X2-CH0-CH-CHn-CHo or X2-CH_-CH-CH-CH_OR5 2 2 || 2 2. | 2 | 2 2 | t 2 0 0 0 0 0 0 \ / \ / \ 'R1 / C \ 12 R ^ R12 R ^ R12 XI11 68054 1 12 in which formulas R is defined in method A, R is 1 7 11 2 1-8 carbon-containing alkoxy or R, R, R and X are as defined in Method C and R is as defined in Method B1, followed by hydrolysis of the protecting groups to form a compound of formula I wherein or R E is hydroxy, optionally converting the resulting base to a pharmaceutically acceptable salt or the resulting salt is converted to a base or another pharmaceutically acceptable salt and optionally the resulting isomeric mixture is separated into the pure enantiomeric isomer A process according to claim 1, characterized in that 9- (3,4-dihydroxybutyl) guanine is prepared. process, characterized in that 9- (2,4-dihydroxybutyl i) guanine. Process according to Claim 1, characterized in that 9- (2,3,4-trihydroxybutyl) guanine is prepared, optionally in erythro or treano form. Process according to Claim 1, characterized in that 9- (3-fluoro-4-hydroxybutyl) guanine is prepared. Process according to one of the preceding claims, characterized in that the compound of the formula I is prepared in the form of its optical isomer. Process according to one of Claims 1 to 6, characterized in that the compound of the formula I is prepared in the form of its physiologically tolerable salt. Process according to Claim 7, characterized in that the compound of the formula I is prepared in the form of its hydrochloride salt. 68054 1. For the preparation of therapeutically active substances with the formula O il N HN I ---- ^ “CH2 ~ CH (R1) CH (R2) CH2OH I color and R2, which are still or or fluorine, for which R1 or R2 is present, R1 and R2 are not present and R1 and R2 are hydroxy or fluorine or R2, or R2 is physiologically acceptable, or if the optical isomer is acceptable, känne-t ecknat av att A. en förening med formeln OT i | ^ HN -N "CH2CH <R1) CHiR2) C00R11 color R1 and R2 is an anhydrous and color R1 is an alkyl, 1-8 cholatomic alkyl, substituted or unsubstituted phenyl or benzyl, reducing, B1. f O KT JN - CH0CH - CH - CHo0R3 HN ^^ N fl I 3 “2 I 6 H OR R OR4 H IV OR4 OR3 HFFH _ FF color Y är OH eller en grupp X1 som genom hydrolys kan omvandlas II till en hydroxigrupp, chlorine, bromine, iodine or the group -SH, -OR2, -SR2 or -SO2R2, i and form alkyl R2 with alkyl of 1-8 cholaters, fluoroalkyl of alkyl with 1-8 cholatomers, alkylaryl with 6805 4 benzyl, or aryl, and R2, R4, R8 and R8 are lychees or olives 2 2 2 and betecnar Väte, -COR or -SO2R, color R is an anhydrous process, hydrolysers, B2 and in the first B1 form as defined in formula IV, 3 4 3 5 color ytterligare R och R tillsammans eller R och R tillsammans 4 5 eller R och R tillsammans bildar en cyclisk ketal som definierats i förfarande E, eller en cyclisk karbonatester, hydrolysers, C. and compounds of formula R7 N'-: '-'N · I I 1 VIII NH 11 1 R condensers of compounds of formula X2 - CH2 - CH - CH - CH.R10 H R8 R9 H R9 R8 H P F H | FF 7 color R is en silyl group as trimethylsilyloxy or tert, - butyldiphenylsilyloxy or group Y is defined as for B1, R 10 and R, R and R are lily or ollycyclic silyl groups or trimethylsilyloxy or tert-butyldiphenylsilyloxy or 1 2 3 groups Z, Z and Z are each other or ollycated halogens, chlorine or iodine OR4 or 5 2 OR as defined in form B1 or in form B2, and X 2 in which the group is selected from chlorine, bromine, iodine or group SO ~ R, 2. color R is defined in form B1, the same as group R ^ -R11 hydrolysers , 68054 D. and for formulations R7 N '' Y- NH2 I \ I NH - CH .. - CH - CH - CH R10 R11 R8- 2 r9 H R9 R8 VII HFFHFF _ 7 8 9 10 11 color R, R , R, R and R are as defined for C, the genome of the ring is determined with myrsyra or 7 11 derivative of the orthoformate ester, group R -R hydrolysers, or E. and compounds of formula R7 XI1 "I * 1 —NH HN R11 condensers of formula X2-CH ~ -CH" -CH - CH- or X2-CH0-CH-CH0-CHo or X2-CH -CH-CH-CH DR5 l 2. \ | 2 2 | 2 | 2 2 | i 2 n 0 0 0 0 0 0 Xc / \ c ^ VIII 1 / s12 1x \ 12 'ix 19 RRRR Δ R1 R12 1 12 i vilka formler R är som definierats i förfarande A, R är 17 11 2 alkoxy med 1- 8 cholatomer or R; R, R and X are as defined for C and R are as defined for B1, which represents a hydrolysis of the group consisting of the compounds of formula 1 to R1 or R2 and hydroxy, optionally